Cai-based systems and methods for the localized treatment of uveitis

ABSTRACT

This invention relates to the treatment of uveitis, in particular posterior infectious uveitis. In specific embodiments, the invention provides for methods of treating uveitis and/or infectious uveitis comprising administration of a sustained-release system containing 5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide) and optionally a glucocorticoid. In one embodiment, the glucocorticoid is dexamethasone. In another embodiment, the sustained-release system comprises a polymer such as e.g. polylactic-coglycolic acid.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.14/884,514, filed Oct. 15, 2015, which claims priority to U.S.Provisional Patent Application No. 62/064,815, filed Oct. 16, 2014, thecontents of which are incorporated herein in their entirety.

This application is related to U.S. patent application Ser. No.14/083,794, filed Nov. 19, 2013, which is a continuation application ofU.S. patent application Ser. No. 13/181,847, filed Jul. 13, 2011 (nowU.S. Pat. No. 8,614,235, issued Dec. 24, 2013), which is a continuationapplication of U.S. patent application Ser. No. 11/235,795 (nowabandoned), filed Sep. 26, 2005, which claims the benefit of U.S.Provisional Patent Application No. 60/612,683, filed Sep. 24, 2004, eachof which are herein incorporated by reference in their entirety.

BACKGROUND OF THE INVENTION

5-amino-1,2,3-triazole-4-carboxamide derivatives were originallydiscovered as antiparasitic agents and then subsequently demonstrated tobe antiproliferative agents and potential cancer therapeutics. Thespecific compound,5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide(Formula I illustrated below, shown as the free base form), has beendemonstrated to have antiproliferative and antimetastatic activity thatwas linked to decrease of intracellular calcium by inhibition ofnon-voltage-gated calcium channels. It is an inhibitor of transmembranecalcium influx and intracellular calcium requiring signalling pathways.Tyrosine kinase and metalloproteinase pharmacological mechanisticactivities and anti-angiogenesis activity relevant to antitumor efficacyhave also been described for this compound. The compound of Formula Iwill be referred to in the following discussion using the acronym CAI(carboxy-amido-triazole) which is generally used to describe thecompound.

Clinical investigations have been conducted with CAI in the treatment oflife-threatening diseases. For example, CAI has been used in thetreatment of a variety of refractory tumors, including prostate cancer,lymphomas, glioblastoma, peritoneal cancer, fallopian tube cancer,epithelial ovarian cancer, advanced renal cell carcinoma, metastaticrenal carcinoma, and non-small cell lung cancer (Bauer, K. S. et al.Clin Cancer Res. 5:2324-2329, 1999; Kohn, E. C. et al. Cancer Res.52:3208-3212, 1992; Kohn, E. C. et al. J Biol Chem. 269:21505-21511,1994; Kohn, E. C. et al. Proc Natl Acad Sci USA. 92:1307-1311, 1995;Kohn, E. C. et al. Cancer Res. 56:569-573, 1996; Kohn, E. C. et al. JClin Oncol. 15:1985-1993, 1997; Kohn, E. C. et al. Clin Cancer Res.7:1600-1609, 2001.

While these studies using CAI indicate that CAI has intrinsic clinicalefficacy for many cancer types, the systemic oral dosage regimens andformulations used in a clinical setting to date have been associatedwith dose limiting side effects. Moreover, toxic effects (cerebellarataxia, peripheral neuropathy and exacerbation of depression) have beenobserved in clinical studies at doses of CAI required to achievecirculating levels that are within a narrow range or those projectedfrom pharmacological studies to be required for effective inhibition ofpathological neovascularization. Furthermore, a serious side effectassociated with clinical use of CAI by systemic administration has beenthe loss of vision for which two cases have been described (Berlin, J.et al. Clin Cancer Res. 8:86-94, 2002). Therefore, use of CAI ascurrently applied in clinical investigations for cancer is effectivelyprecluded for acute or chronic treatment of non-life threateningconditions, in particular for the treatment of ocular diseases describedherein.

To treat certain ocular diseases, such as age-related maculardegeneration (or ARMD), and diabetic retinopathy, or diseases withspecific ocular manifestations, such as von Hippel landau syndrome,therapeutic treatments rely on occlusion of the blood vessels usingeither threshold laser photocoagulation, or 0 laser combined with aphotoactivated dye. However, such treatment requires eitherfull-thickness retinal damage by thermal destruction, or damage tomedium and large choroidal vessels thereby precluding any potentialvisual recovery. Further, the subject is left with a scar and visualscotoma. Moreover, recurrences are common, and visual prognosis is poor.

Recent research in the treatment of neovascularization has had the aimof causing a more selective closure of the blood vessels, in order topreserve the overlying neurosensory retina. Such strategies have beenused for the treatment of diabetic retinopathy, the leading cause ofblindness among working age adults in Europe and the United States.However, extensive ocular tissue damage can occur after panretinalphotocoagulation, with the visual handicap of more limited peripheralvision and poor night vision. With focal laser treatment,photocoagulation often can further compromise macular blood flow.Alternatively, a variety of molecules are in development or have beenapproved that target angiogenic pathways (e.g. the VEGF pathway). Thus,using antiangiogenic compounds is an alternative to lasering ofpatients.

CAI is an anti-angiogenic and anti-parasitic compound; however, the pooraqueous solubilities of CAI compounds, as well as reported neurotoxicityfor CAI, means that novel methods of administration and targetedadministration of CAI compounds are required for providing safe andeffective doses to treat disease and non-life threatening diseases inparticular.

High local concentrations of CAI may be required to treat acute diseasesymptoms while lower concentrations can be effective as continuationtherapy or prophylactic therapy. Additionally the frequency ofadministration of a formulation of a CAI compound can also be used toensure safe and effective local concentrations to slow vascularoutgrowth. Treatment may be necessary from every week, to every month tofew months, to yearly dosing with appropriate molecules in sustaineddelivery systems.

Posterior segment neovascularization (NV) is the vision-threateningpathology responsible for the two most common causes of acquiredblindness in developed countries: exudative ARMD and proliferativediabetic retinopathy (PDR). Currently the only approved treatments forposterior segment NV that occurs during exudative ARMD are laserphotocoagulation, photodynamic therapy with VISUDYNE®, and the VEGFbinding oligonucleotide aptamer Macugen®. Laser and photodynamictherapies involve occlusion of affected vasculature, which results inlocalized laser-induced damage to the macula. For patients with PDR,surgical interventions with panretinal laser, or vitrectomy and removalof preretinal membranes, or treatment with Macugen® are the onlytreatment options currently available. However, in addition to therecently approved anti-VEGF aptamer oligonucleotide Macugen®; severaldifferent compounds are being evaluated clinically, including, forexample, anecortave acetate (Alcon Research, Ltd.), and Lucentis®(ranibizumab, Genentech), Squalamine lactate (Genaera Corporation),LY333531 (Lilly), Cand5 (Acuity Pharmaceuticals), Talaporfin sodium(Light Sciences Corp.), and Fluocinolone (Bausch & Lomb).

Treatments using dosage regimens, routes of administration andformulations of CAI described to date do not have adequate safety fortreating severe proliferative diseases that are non-life threatening.There exists an unfulfilled need for new dosage regimens, routes ofadministration and formulations of CAI that can provide therapeuticeffects on non-life threatening proliferative diseases, as exemplifiedby ocular diseases that are characterized by neovascularization andpathological cellular proliferation and invasion. There also exits anunfilled need for administering CAI formulations that can providetherapeutic effects on uveitis, which is characterized by the swellingof the middle layer of the eye and which according to Barisani-Asenbaueret al. “is the fourth most common cause of blindness among theworking-age population in the developed world.” Orphanet Journal of RareDiseases 2012, 7:57.

BRIEF SUMMARY OF THE INVENTION

The subject invention provides novel sterile aqueous formulations of CAIcompounds, their use in the localized treatment of ocular diseases, aswell as methods for their manufacture. In some embodiments of theinvention, novel and advantageous CAI compounds, their respectiveformulations, and methods for their use are provided. The novel CAIcompounds and aqueous, sterile formulations of the invention areparticularly useful in the localized treatment of non-life threateningdiseases such as proliferative diseases, ocular diseases or conditions,inflammatory conditions, edematous diseases, signaltransduction-mediated diseases, and matrix metalloproteinase-mediateddiseases, and neovascular diseases. In particular, the novel CAIcompounds and aqueous, sterile formulations of the invention when usedin a sustained release system are useful for the treatment of uveitisand especially posterior infectious uveitis.

In certain embodiments, CAI compounds of the present invention arecrystalline and small particulate pure free base forms of CAI, CAIprodrugs, and CAI acid addition salt forms. Further embodiments of theinvention provide novel, therapeutically effective formulations ofcrystalline and small particulate pure free base forms of CAI compoundsof the invention. In other embodiments, the CAI compounds of the claimedinvention are provided as sonicated microparticles or in a complex, suchas e.g. in complex with beta-cyclodextrin. When formulated asmicroparticles or complex, the CAI compounds may be further used withtime-release delivery systems.

Specific diseases that can be treated with the CAI compounds of theinvention include, but are not limited to, ocular disease includingneovascular ocular disease, edematous ocular disease, ocular tumors, andintraocular inflammation; dermatological disease including psoriasis,eczema, actinic keratosis, and rosacea; inflammatory disease includingarthritis and arthrosis; neurological diseases includingneurodegenerative disease, pain, and epilepsy; cancers including cancersof the bladder, brain, breast, colon, endometria, kidney, lungs,pancreas, and thyroid; and proliferative vascular disorders includingrestenosis or proliferation associated with angioplasty. Such diseasesinclude, but are not limited to, uveitis (anterior uveitis, intermediateuveitis, posterior uveitis and diffuse uveitis). In one embodiment, theCAI compounds of the invention may be used to treat posterior uveitis,such e.g. posterior infectious uveitis. In yet another embodiment, theCAI compounds of the invention may be used to treat posterior infectiousuveitis caused by toxoplasmosis, toxocara and visceral larva migrans orother parasitic diseases.

Aqueous, sterile formulations as described herein provide for thelocalized administration of high concentrations of CAI compounds (whichare normally poorly soluble) for the treatment of such acute therapeuticindications as exemplified by severe neovascular ocular disease stagesof “wet” ARMD or diabetic retinopathy. Similarly, for chronic use, thesubject invention provides sterile, aqueous formulations for thecontrolled release of CAI compounds via oral, systemic or local routesof administration, where such formulations ensure the maintenance oftherapeutic CAI compound drug levels with low variability andreliability for chronic application.

Treating localized proliferative diseases, including those characterizedby neovascular pathology, by the local administration of CAI compoundsof the invention as described herein, provides significant advantages tothe systemic administration of CAI with such systemic administrationhaving been practiced in cancer to date.

Local administration of CAI compounds as provided herein allows for themaintenance of efficacious, non-toxic levels of active CAI free drug atthe local tissue disease site, thus providing a high margin of safetyand enables therapeutically effective treatment of diseases,particularly non-life threatening diseases. The localized treatment ofnon-life threatening diseases using CAI compounds as described hereinprovides significant advantages relative to systemic treatment withforms and formulations of CAI described to date, which have unacceptablelow safety thresholds.

Similarly, according to the subject invention, the local administrationof CAI compounds, and formulations thereof, by means of a drug deliverydevice or implant placed in proximity to the local tissue site providesfor the maintenance of efficacious, safe levels of active CAI drugingredient at the local tissue disease site. Thus, the subject inventionprovides a high margin of safety and enables use in treating non-lifethreatening diseases for which CAI dosed systemically, in formsdescribed to date, is practically precluded.

In one embodiment, local administration of CAI compounds as describedherein is suitable for acute disease therapy and induction of remissionof disease signs and symptoms. In another embodiment, localadministration of CAI compounds as described herein is suitable forchronic disease therapy and maintenance of remission of disease signsand symptoms.

According to the subject invention, the local ocular administration ofCAI compounds of the invention, and/or formulations thereof, attenuateocular pathological disease processes without unduly compromising normalhealthy ocular function. Thus, local ocular administration of a CAIcompound of the invention, and/or formulations thereof, provides for anefficacious but safe controlled concentration range of CAI directly inthe eye.

Ocular CAI-based therapies, as describe herein, provide significantadvantages for treating neovascular ocular disease relative to currentlaser surgery treatment modalities including panretinalphotocoagulation, which can be accompanied by extensive ocular tissuedamage. In the examples of posterior neovascular ocular diseases, suchas Age-related Macular Degeneration and Diabetic Retinopathy, targetocular pathologies and tissues for treatment are especially localized tothe retinal, choroidal and corneal ocular compartments.

It is contemplated herein that CAI compounds of the invention canreadily penetrate the human scleral tissue after periocularadministration in patients with ocular disease. This particular propertyof CAI compounds and CAI-based formulations of the invention enableseffective minimally invasive delivery of the parent CAI therapeuticentity (or also referred to herein as CAI free base) to the patient'sdiseased ocular tissue compartment(s) in therapeutic concentrationwithout causing ocular toxicity that has been previously observed withthe systemic administration of the biologically active CAI parent drugto cancer patients.

According to the subject invention, tissue targets for localizedtreatment using the CAI compounds include, but are not limited to,dermal, ocular, pulmonary, vascular, joint, nasal, ictic, bone,gastrointestinal, and localized neural tissues e.g. brain and spinalcolumn of the central nervous system. Accordingly, administrations ofCAI compounds of the invention as described herein can be used to treatocular, dermatological, pulmonary, vascular, joint, bone, nasal, ictic,gastrointestinal and neurological diseases. In a one embodiment, CAIcompounds of the invention, and/or formulations thereof, are useful fortreating local disease pathologies that are targeted by the establishedpharmacological mechanism(s) of action of CAI to inhibit thepathological increase in intracellular calcium concentration that blockshyperproliferative, inflammatory, neurodegenerative, edematous,macroscopic pathologies associated with localized target diseasesdescribed herein. In another embodiment, the CAI compounds of theinvention, and/or formulations thereof, are useful for treating localdisease pathologies associated with inflammation due to the presence ofparasites (such as e.g. Toxoplasma gondii) in the eye.

In an alternate embodiment, the present invention provides CAI compoundsand CAI compound formulations that provide for controlled release ofpharmacologically active CAI in a targeted tissue and at therapeuticallyeffective concentrations. Such CAI compounds can be locally delivered oradministered to a target tissue in vivo by passive spontaneous,biologically accelerated, or assisted (such as sonophoresis or galvanic,e.g. iontophoresis) mechanisms.

The CAI compounds of the present invention and formulations thereof, areadvantageous because they overcome problems associated with sterility,stability, toxicity, lack of target tissue specificity, safety,efficacy, extent, and variability of bioavailability, which exist withthe administration of the active form of CAI. The CAI compounds of thepresent invention, and CAI compound formulations, are particularlyadvantageous in minimizing the undesirable side effects associated withcurrent CAI treatment or therapy, such as loss of vision, cerebellarataxia, peripheral neurotropathy, and exacerbation of depression.

As contemplated in the subject invention, where a CAI compound comprisesa CAI prodrug, the CAI prodrug can be converted to a biologically activeCAI compound at a controlled rate via passive (such as by aqueoushydrolysis) or biologically mediated (such as biocatalytic or enzymatic,e.g. by intraocular esterases) mechanisms. An advantage of the in vivoconversion of the CAI-based prodrug to CAI is that the ensuing CAIprovides localized therapeutic effects in target disease tissue withhigh therapeutic margins of safety.

A further embodiment provides the use of CAI compounds in conjunctionwith a drug delivery system in the form of an implant or a device forthe treatment of conditions as set forth herein. Certain embodiments ofthe invention contemplate the use of CAI compounds and formulationsthereof for use comprising a coating for example of a device inconjunction with physical device implants such as stents and bandligatures. Therapeutic uses of such implants include but are not limitedto vascular diseases such as restenosis, and in bone and tissue grafts.

A further embodiment of the subject invention provides for the localadministration of CAI compounds in combination with otherpharmacological therapies. As contemplated in the subject invention,combination therapies of CAI compounds with other medicaments targetingsimilar or distinct disease mechanisms have advantages, which includegreater efficacy in therapeutic treatment of a disease and greatermargins of safety relative to respective monotherapies with eitherspecific or separate medicament.

In one embodiment, a CAI compound is used in a method to treatneovascular ocular disease comprising localized (for example, in oculartissue) concurrent administration with one or more other medicamentseach of which may act to block angiogenesis by a pharmacologicalmechanism. Such combinations would act by providing for superiorefficacy from mechanisms of action from additivity, synergy, oramelioration of side effects. Medicaments that can be concurrentlyadministered with a CAI compound of the invention include, but are notlimited to, vascular endothelial growth factor VEGF blockers (e.g. byVEGF neutralizing binding molecules such as Macugen® (Eyetech) andLucentis® (ranibizumab, Genentech), Squalamine lactate (GenaeraCorporation); and VEGF tyrosine kinase inhibition) for treatingneovascular ocular disease (ARMD and Diabetic Retinopathy) andglucocorticoids (e.g. Triamcinolone) for treating macular edema. Othergrowth factors that can be concurrently administered with the CAIcompounds of the invention (preferably in sterile, aqueous formulations)include, but are not limited to, Pigment Epithelium Derived Factor(PEDF), Erythropoietin (EPO), integrin, endostatin, R-cadherin,angiostatin, and fibroblastic growth factor.

The systems and method may include treatment of uveitis with CAIcompounds alone or as part of combination therapy. In particular, incertain embodiments of the invention, CAI may be used in a time-releaseformulation to treat posterior infectious uveitis.

In certain embodiments, the invention provides for using a CAI compoundsuch as e.g.5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide)in the treatment of uveitis, in particular posterior infectious uveitis.When used to treat uveitis (e.g. posterior infectious uveitis), the CAIcompound may be used in conjunction with any of the other optionalcomponents described herein such as antibacterial agents and/oranti-inflammatory agents. In certain embodiments, the CAI compound aloneor conjunction with other components is administered in a time-releaseformulation such as e.g. a sustained release system. Accordingly, oneembodiment of the invention is a method of treating uveitis includingadministering a sustained-release system containing a pharmaceuticallyeffective amount of5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide)and optionally a glucocorticoid to a patient. Thus, in one embodiment ofthe invention, the method includes administration of a sustained-releasesystem containing a pharmaceutically effective amount of5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide).In other embodiments, the method includes administering asustained-release system containing a pharmaceutically effective amountof5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide)and a glucocorticoid to a patient. The method may also includediagnosing uveitis in the patient. In certain embodiments, thesustained-release formulation may include a polymer such as e.g.polylactic-coglycolic acid (PLGA). In one embodiment, the polymer ispoly (D,L-lactide-co-glycolide) PLGA.

While a variety of glucocorticoids may be used, in one embodiment, theglucocorticoid may be dexamethasone such as e.g. from about 0.5 to about0.9 mg of dexamethasone. The sustained-release system may includeadditional components including an anti-inflammatory agent and/or anantibacterial agent. The sustained-release system may be administered byintravitreal injection or intravitreal implantation. In one embodiment,the sustained-release system includes poly (D,L-lactide-co-glycolide)PLGA and dexamethasone. In one particular embodiment, thesustained-release system includes an OZURDEX® (dexamethasone)intravitreal implant.

In another embodiment, the invention provides for methods of treatinginfectious uveitis. The method includes administering asustained-release system containing a pharmaceutically effective amountof5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide)and optionally glucocorticoid to a patient. Accordingly, in oneembodiment, the method includes administering a sustained-release systemcontaining a pharmaceutically effective amount of5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide)and glucocorticoid to a patient. The method may also include diagnosingposterior infectious uveitis in the patient. In certain embodiments, thesustained-release formulation includes a polymer. In one embodiment, thepolymer contains polylactic-coglycolic acid (PLGA). In one specificembodiment, the polymer is poly (D,L-lactide-co-glycolide) PLGA. Incertain embodiments, in methods in which a glucocorticoid is used, theglucocorticoid includes or is dexamethasone. In certain embodiments, thesustained-release system contains from about 0.5 to about 0.9 mg ofdexamethasone. In other embodiments, the sustained release systemcontains from about 150 mcg to about 200 mcg of5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide).The sustained-release system may contain a variety of additionalcomponents. In one embodiment, the sustained-release system include ananti-inflammatory agent and/or an antibacterial agent. The method mayinclude various routes of administration. In certain embodiments, themethods include intravitreal injection or implantation. In oneembodiment, the method for treating posterior infectious uveitisincludes administration of a sustained-release system containing poly(D,L-lactide-co-glycolide) PLGA, dexamethasone and5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide).The sustained-release system may be an implant. In one particularembodiment, the sustained-release system is an OZURDEX® (dexamethasone)intravitreal implant to which5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide)has been added.

The invention also generally provides for methods of treating uveitis byadministering a sustained-release system containing a pharmaceuticallyeffective amount an anti-parasitic or anti-bacterial agent and aglucocorticoid to a patient. The method may also include diagnosinguveitis in the patient. The sustained-release formulation may contain apolymer. In certain embodiments, the polymer includespolylactic-coglycolic acid (PLGA). In one embodiment, the polymerincludes poly (D,L-lactide-co-glycolide) PLGA. In one embodiment, theuveitis is posterior infectious uveitis. The anti-parasitic agent may be5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide).The glucocorticoid may be dexamethasone. In some embodiments, thesustained-release system contains from about 0.5 to about 0.9 mg ofdexamethasone. The sustained-release system may also further contain ananti-inflammatory agent. The step of administration may includeintravitreal injection and/or implantation. In one embodiment, thesustained-release system includes poly (D,L-lactide-co-glycolide) PLGAand dexamethasone. Thus, the sustained-release system may include anOZURDEX® (dexamethasone) intravitreal implant.

In one embodiment, the invention provides for a method of treatinguveitis in a patient by administering a sustained-release systemcomprising a pharmaceutically effective amount of5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide)(CAI). In one embodiment, the uveitis is posterior infectious uveitis,which in certain embodiments may caused by toxoplasmosis, toxocara orvisceral larva migrans. The sustained release-system may be a polymer,which may degrade over time. In certain embodiments, the polymer may bepolylactic-coglycolic acid (PLGA) such as poly(D,L-lactide-co-glycolide) PLGA. In other embodiments, the sustainedrelease system may be a non-erodible intravetreal implant. Thesustained-release system further includes one or more of ananti-inflammatory agent, a steroid and/or an antibacterial agent. Incertain embodiments, the steroid is a glucorticoid (such as e.g.dexamethasone) or fluocinolone acetonide. In one embodiment, thesustained-release system may from about 0.5 to about 0.9 mg ofdexamethasone. The sustained release system may administered by avariety of routes including but not limited to intravitreal injectionand intravitreal implantation. In one embodiment, the sustained-releasesystem contains poly (D,L-lactide-co-glycolide) PLGA and dexamethasone.The sustained-release system is an implant. When used as part of asustained release system, CAI may be provided in a variety of differentformulations. For example, CAI may be provided as a sonicatedmicroparticle or in a molecular complex in formulation withhydroxypropyl β-cyclodextrin. In certain embodiments, CAI is in amolecular complex formulation with 1 to 60 wt % hydroxypropylβ-cyclodextrin and one or more of glutamate, L-phenylalanine, acellulose derivative (such as e.g. hydroxypropyl methyl cellulose), anda surfactant. The sonicated CAI microparticle or CAI molecular complexmay have a diameter of less than 10 micrometers. The sustained-releasesystem may include an OZURDEX® (dexamethasone) intravitreal implant oran ILUVIEN® (fluocinolone acetonide) intravitreal implant.

Another embodiment of the invention is a method of treating posteriorinfectious uveitis (such as e.g. posterior infectious uveitis caused bytoxoplasmosis, toxocara or visceral larva migrans) in a patientcomprising administering a sustained-release system comprising apharmaceutically effective amount of5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide)and a steroid to a patient. In certain embodiments, the steroid is aglucocorticoid or fluocinolone acetonide. The sustained-release systemmay be a polymer such as for example, polylactic-coglycolic acid (PLGA)(e.g. poly (D,L-lactide-co-glycolide) PLGA). The polymer may degradeover time. The steroid may be dexamethasone. In certain embodiments, thesustained-release system contains from about 0.5 to about 0.9 mg ofdexamethasone. In other embodiments, the sustained-release system alsocontains an anti-inflammatory agent and/or an anti-bacterial agent. Thesustained release system may be administered by intravitreal injectionor implantation. In certain embodiments, the sustained-release systemcontains poly (D,L-lactide-co-glycolide) PLGA and dexamethasone. Inother embodiments, the sustained-release system is an implant. The CAImay be provided a sonicated microparticle or in a molecular complex informulation with hydroxypropyl β-cyclodextrin. The sonicated CAImicroparticle or molecular complex may have a diameter of less than 10micrometers. In certain embodiments, CAI is in a molecular complexformulation with 1 to 60 wt % hydroxypropyl β-cyclodextrin and one ormore of glutamate, L-phenylalanine, a cellulose derivative (e.g.hydroxypropyl methyl cellulose), and a surfactant. In one embodiment,the sustained release system may be a non-erodible intravitreal implant.The sustained-release system may include an OZURDEX® (dexamethasone)intravitreal implant or an ILUVIEN® (fluocinolone acetonide)intravitreal implant.

The invention encompasses the use of any of the CAI compounds andformulations thereof in the manufacture of a medicament for the localtreatment of a non-life threatening disease. In one embodiment, theinvention encompasses the use of a CAI compound and a formulation, andpreferably an aqueous formulation, thereof in the manufacture of amedicament for the local treatment of an ocular disease.

Other features and advantages of the invention will be apparent from thedetailed description and examples that follow.

DETAILED DESCRIPTION OF THE INVENTION

Uveitis is inflammation or swelling of the uvea—the middle layer of theeye that consists of the iris, ciliary body and choroid. “Because thedisease involves recurrent intraocular inflammation, uveitis can causetransient or permanent visual impairment and ocular complication thatare not responsive to therapy.” Barisani-Asenbauer et al., OrphanetJournal of Rare Diseases 2012, 7:57, the disclosure of which isincorporated as it pertains to uveitis. There are different types ofuveitis based on where the inflammation occurs in the uvea:

Anterior uveitis is inflammation of the iris (iritis) or the iris andciliary body;

Intermediate uveitis is inflammation of the ciliary body;

Posterior uveitis is inflammation of the choroid; and

Diffuse uveitis (also called panuveitis) is inflammation of all areas ofthe uvea.

The cause of uveitis is often unknown and frequently occurs in otherwisehealthy people. However, it can also be associated with another illnesssuch as an autoimmune disorder or an infection from a virus or bacteria.According to Barisani-Asenbauer et al. almost 20% of all uveitis casesinvolve infectious agents with ocular toxoplasmosis account for 7% ofall cases and herpetic uveitis accounting for 7.1% of all cases.Barisani-Asenbauer et al. at pg. 5. In particular, causes of posterioruveitis include, but are not limited to, syphilis, toxoplasmosis,tuberculosis, endogenous endophthalmitis, and viral causes (includingherpes simplex virus, herpes zoster virus, and cytomegalovirus). “Inposterior uveitis cases 29% were caused by ocular toxoplasmosis and17.7% by multifocal choroiditis.” Barisani-Asenbauer et al. at Abstract.Common systemic infectious causes of uveitis include but are not limitedto Mycobacterium tuberculosis, Ascaris lumbricoides, Candida albicans,Chlamydia trachomatis, Filaria, Aspergillus fumigatus, Treponemapallidum, Giarda lamblia, Borrelia Burgdorferi, and Plasmodiumfalciparum. Barisani-Asenbauer et al. Exemplary parasitic agents whichcause uveits include but are not limited to Toxoplasma, Toxocara(Oxyuris or ascariasis), and larva parasites. BenEzra et al., Br JOphthalmol., 2005; 89(4): 444-448, the disclosure of which isincorporated as it pertains to uveitis. A variety of symptoms may occuras a result of suffering from uveitis including blurred vision, floaters(dark floating spots in the field of vision), and eye pain. There arecurrently various treatments of uveitis. Most commonly, most types ofuveitis including posterior uveitis are treated by the administration ofsteroids either orally or as eye drops. However, for infectiousposterior uveitis, treatment with steroids does not address theunderlying cause of the infection—the parasite. As such, there exists aneed for a treatment for uveitis in particular infectious uveitis thatnot only relies on common steroid treatment but also eliminates theunderlying cause of the inflammation.

According to the Merck Manual, toxoplasmosis (infection with Toxoplasmagondii) is one of the most common causes of retinitis (includingposterior infectious uveitis) in immunocompotent patients. For posteriorinfectious uveitis caused by toxoplasmosis, the symptoms of floaters anddecreased vision may be due to cells in the vitreous humor or to retinallesions or scars. Current treatments involve multidrug therapy includingpyrimethamine, sulfonamides, clindamycin, and, in select cases, systemiccorticosteroids. See Merck Manual, Uveitis and related disorders.

The subject invention provides CAI compounds and formulations thereof,and methods for their use in the localized treatment of non-lifethreatening diseases. Formulations of CAI compounds of the subjectinvention can comprise CAI as a free base and a CAI prodrug inmicrocrystalline form, as a microparticle, as an emulsion, and the like.The subject invention further provides methods for treating non-lifethreatening diseases comprising administration to a patient of atherapeutically effective amount of a CAI compound of the invention(i.e., using novel delivery systems and combination therapies), that areeffective and are associated with little or no adverse side effects.

In one embodiment, the invention provides for CAI compounds intime-release formulations, which may utilize CAI's anti-parasitic and/oranti-proliferative properties. In particular, the invention provides forCAI compounds in a time-release formulation to treat uveitis especiallyposterior infectious uveitis. When the CAI compound of the claimedinvention are used in such a time-release formulation, the time-releaseformulation may include other therapeutic compounds such as ananti-inflammatory agent, an antibacterial agent or a glucorticoid. Thus,the invention provides for the use of CAI compound in combinationtherapy for the treatment of uveitis especially posterior infectiousuveitis. Moreover, the invention provides for methods of reducing theinflammation in the eye due to the presence of a parasite (such as e.g.Toxoplasma gondii) by administering CAI compound in a time-releaseformulation.

Routes of administration contemplated herein for the delivery of CAIcompounds and/or of formulations thereof by local administrationinclude, but are not limited to, ocular, dermal, nasal, ictic,pulmonary, intravitreal, peribulbar, subtenon, periocular, retrobulbar,subretinal, and posterior juxtascleral subconjunctival routes. Suchroutes of administration may be performed by injection by syringe andusing a needle or cannula or by needle-free systems: topicaladministration via a formulation comprising drops, ointment, and creams,inhalation by means of an inhalation device such as a metered doseinhaler or dry powder inhaler or nebulizer, or in conjunction with drugdelivery systems exemplified by controlled release of a CAI compoundfrom a matrix comprising a contact lenses (for delivery of drug to atissue of the eye), from a device, from an implant, for release of drugto proximal tissue, and by use of an iontophoretic system e.g., toenhance the rate of penetration of drug through a barrier tissue.

In an embodiment of this invention, a CAI compound, or formulationthereof, is administered locally to ocular tissue via a route thatattenuates ocular pathological disease processes without undulycompromising normal or healthy ocular function by acting directly in theeye (e.g., at the site of diseased tissue) in a controlledtherapeutically effective and localized fashion.

In particular embodiments, the invention provides for using CAIcompounds (such as e.g.5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide))in the treatment of uveitis (including posterior infectious uveitis).The CAI compounds may be used alone or in combination with othercomponents. In certain embodiments, the invention provides for thetreatment of uveitis (in particular posterior infectious uveitis) usinga sustained-release system (i.e. a time-release system). Thistime-release system may be a polymeric implant containing the CAIcompound and additional agents. In certain embodiments, the polymericimplant also includes a glucocorticoid such as e.g. dexamethasone. Inone embodiment, the sustained-release system may be an OZURDEX®(dexamethasone) intravitreal implant containing the CAI compound (e.g.5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide)).

I. DEFINITION OF TERMS

The term “CAI compound,” as used herein, refers to CAI as a free baseand to CAI analogs, and CAI prodrugs, and to any salts such as acidsalts thereof. CAI (carboxy-amido-triazole) has the following structure:

The CAI compounds of the invention are particularly useful in thetreatment of non-life threatening diseases.

The term “therapeutically effective amount,” as used herein, refers tothe amount necessary to elicit the desired biological response such asamelioration or reduction in severity of a symptom by a percentageamount. In accordance with the subject invention, the therapeuticallyeffective amount of a CAI compound is the amount necessary to treatnon-life threatening diseases. In one embodiment, an effective amount ofa CAI compound, or formulation thereof, may ameliorate the severity ofsymptoms and/or complications associated with a non-life threateningdisease. The amelioration in symptom and/or complication severity may bea 5%, 10%, 15%, 20%, 25% 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,75%, 80%, 85%, 90%, 95%, 98% or 99% decrease in severity. Preferably,the therapeutically effective amount of CAI compound locallyadministered in the treatment of ocular disease is a dose of 5 mg to 50mg. More preferably, 5 mg is dosed intravitreally and 50 mg is dosedperiocularly. More preferably, the CAI compound is CAI free base.

The term “patient,” as used herein, describes an organism, includingmammals, to which treatment with the compositions according to thepresent invention is provided. Mammalian species that benefit from thedisclosed compounds and methods of treatment include, but are notlimited to, apes, chimpanzees, orangutans, humans, monkeys; anddomesticated animals (i.e., pets) such as horses, dogs, cats, mice,rats, guinea pigs, and hamsters.

The term “about” when referring to a measurable value such as an amount,a temporal duration, and the like, is meant to encompass variations of±20% or ±10%, more preferably ±5%, even more preferably ±1%, and stillmore preferably ±0.1% from the specified value, as such variations areappropriate to perform the disclosed methods.

The term “treatment” as used herein covers any treatment of a non-lifethreatening disease in a patient, particularly a human, comprisingadministration of a compound and/or formulation of a compound of thisinvention and methods of the subject invention, and includes:

-   -   1. Preventing the disease from occurring in a patient that may        be predisposed to the disease but has not yet been diagnosed as        having it;    -   2. Inhibiting the disease, e.g., inhibiting occurrence of an        additional disease; or arresting development, inducing        remission, or maintaining remission of the disease;    -   3. Relieving the disease, e.g., causing regression of the        disease or addressing by diminishing or reducing symptoms; or    -   4. Inhibiting recurrence of a disease.

“Concurrent administration” and “concurrently administering,” as usedherein, includes administering a compound or formulation in atherapeutic method suitable for use with the compounds and methods ofthe invention (for example, administration of a CAI prodrug of FormulaeII or III, below) in the treatment of serious, non-life threateningdiseases.

“Non-life threatening disease(s),” as used herein, includes but is notlimited to non-life threatening proliferative, inflammatory,neovascular, ocular, edematous, signal transduction-mediated diseases,matrix metalloproteinase-mediated diseases, and neurodegenerativediseases. Examples of serious, non-life threatening ocular diseasesinclude, but are not limited to diabetic retinopathy (DR), neovascularage-related macular degeneration (ARIVID), diabetic macular edema (DME),cystoid macular edema (CME) and ocular tumors such as retinoblastoma(RB), Retinopathy of Prematurity (ROP), Retinal Vascular Occlusions(RVO), corneal neovascularization, iris neovascularization, neovascularglaucoma, ischemic neural damage, uveitis (including, anterior uveitis,intermediate uveitis, posterior uveitis, posterior infectious uveitis,and diffuse uveitis), glaucoma, and pterygium, neovascular diseases ofthe retina such as hyperproliferative retinopathies,vitreoretinopathies, and retinal degeneration associated with systemicdiseases such as diabetes mellitus, ischemic and hypoxic conditionsassociated with retinal vein and artery occlusion (e.g., from sicklecell disease or thrombosis), retinal degeneration resulting from retinaldetachment, and age-related macular degeneration. Non-life threateningdiseases, as defined herein, include any disease that is directly orindirectly mediated by CAI. Non-life threatening diseases are notdiseases that may cause death in a patient. A non-life threateningdisease may be present in a patient together with a life-threateningdisease such as diabetes mellitus. In particular embodiments, thenon-life threatening disease is uveitis or even more preferablyposterior infectious uveitis.

The present invention may also be used to treat ocular symptomsresulting from diseases or conditions that have both ocular andnon-ocular symptoms. Some examples include AIDS-related disorders suchas cytomegalovirus retinitis and disorders of the vitreous; vonHippel-Lindau Syndrome (a disease that has ocular and nonocularneovascularizations); pregnancy-related disorders such as hypertensivechanges in the retina; and ocular effects of various infectious diseasessuch as tuberculosis, syphilis, lyme disease, parasitic disease,toxocara canis, ophthalmonyiasis, cyst cercosis, and fungal infections.Examples of non-ocular diseases include but are not limited torheumatoid arthritis, psoriasis, contact dermatitis, keratitis,conjunctivitis, scleritis, squamous cell carcinoma, condyloma, eczema,rosacea, vascular proliferation associated with angioplasty, graft vshost disease (organ and tissue transplantation), glioblastoma,peripheral neuropathies, diabetic neuropathy, collagen vasculidities.

In addition, the present invention can be used to treat diseases otherthan non-life threatening diseases such as, but not limited to, bladdercancer, breast cancer, brain tumors, colon and rectal cancer,endometrial cancer, kidney cancer, leukemia, lung cancer, melanoma,non-Hodgkin's lymphoma, pancreatic cancer, prostate cancer, skin(non-melanoma) cancer, and thyroid cancer.

The following definitions are applicable to those structures as definedfor Formula II and Formula III, below.

Unless otherwise specified, as used herein, the term “alkyl” refers to astraight or branched chain alkyl moiety. In one embodiment, the alkylmoiety is C₁₋₈ alkyl, which refers to an alkyl moiety having from one toeight carbon atoms, including for example, methyl, ethyl, propyl,isopropyl, butyl, tert-butyl, pentyl, hexyl, and octyl.

The term “alkyl” also includes cycloalkyl, including for examplecyclohexyl and the like. The term “alkyl” also includes alkenyl, whichrefers to a straight or branched alkyl moiety having one or more carbondouble bonds, of either E or Z stereochemistry where applicable, andincludes for example, vinyl, 1-propenyl, 1- and 2-butenyl, and2-methyl-2-propenyl.

The term “alkyl” also refers to an alicyclic moiety having from three tosix carbon atoms and having in addition one double bond. Such groups,also known as “cycloalkenyl,” include, for example, cyclopentenyl andcyclohexenyl.

The term “alkyl” also includes alkynyl moieties, including for example,ethynyl, 1-propynyl, 1- and 2-butynyl, 1-methyl-2-butynyl, and the like.

The term “alkoxy” refers to an alkyl-O-group, in which the alkyl groupis as described herein.

The term “halogen” is refers to fluorine, chlorine, bromine and iodine.

The term “aryl” refers to an aromatic carbocyclic ring, e.g., phenyl,substituted phenyl, and like groups, as well as rings, which are fused,e.g., naphthyl, biphenyl indaryl and the like. Aryl groups may beoptionally substituted with from 1 to 3 groups of alkyl, aryl,heteroaryl, halogen, hydroxy, alkoxy, nitro, cyano, trifluoromethyl,alkylthio, alkylsulfonyl, arylsulfonyl, N(R)₂, S(O)₂N(R)₂, CH₂N(R)₂,CO₂R, and C(O)N(R)₂ wherein R groups are independently hydrogen, alkyl,alkoxyalkyl, hydroxyalkyl, aminoalkyl, arylalkyl, aryl, heterocycloalkylor heteroaryl.

The term “arylalkyl” refers to a moiety in which the “aryl” and “alkyl”groups are as described herein.

The term “heterocycloalkyl” refers to a saturated heterocyclic moietyhaving from two to six carbon atoms and one or more heteroatom from thegroup N, O, and S (or oxidized versions thereof) which may be optionallybenzofused at any available position. This includes, for example,azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, benzodioxolyland the like.

The term “heterocycloalkyl” also refers to an alicyclic moiety havingfrom three to six carbon atoms and one or more heteroatoms from thegroup N, O, and S and having in addition one double bond. Such moietiesmay also be referred to as “heterocycloalkenyl” and includes, forexample, dihydropyranyl, and the like.

In the case that the heterocyclyl moiety contains a nitrogen atom thatmay be substituted, then a substituent may be chosen from the group ofR.

The term “heteroaryl” refers to monocyclic or bicyclic aromatic ringsystems of five to ten atoms of which at least one atom is selected fromO, N, and S, in which a carbon or nitrogen atom is the point ofattachment, and in which one additional carbon atom is optionallyreplaced with a heteroatom selected from 0 or S, and in which from 1 to3 additional carbon atoms are replaced by nitrogen heteroatoms. Theheteroaryl group is optionally substituted with up to three groupsselected from alkyl, aryl, heteroaryl, halogen, hydroxy, alkoxy, nitro,cyano, trifluoromethyl, alkylthio, alkyl sulfonyl, aryl sulfonyl, N(R)₂,S(O₂)N(R)₂, CH₂N(R)₂, CO₂R, and C(O)N(R)₂ wherein R groups areindependently hydrogen, alkyl, alkoxyalkyl, hydroxyalkyl, aminoalkyl,arylalkyl, aryl, heterocycloalkyl or heteroaryl. This term includes, forexample, imidazolyl, tetrazolyl, furanyl, thiophenyl, pyridyl, indolyl,quinolyl, benzothiadiazolyl, benzofurazanyl, benzotriazolyl, and thelike.

Certain of the above-defined terms may occur more than once in FormulasII and III and upon such occurrence each term shall be definedindependently of the other.

“Optional” or “optionally” means that the subsequently described event,element, or circumstance may or may not occur, and that the descriptionincludes instances where said event occurs and instances in which itdoes not.

The term “substituted” shall be deemed to include multiple degrees ofsubstitution by a named substituent.

Where multiple substituent moieties are disclosed, the substitutedcompound can be independently substituted by one or more of thedisclosed or claimed substituent moieties, singly or plurally.

II. CAI PRODRUGS OF THE INVENTION

In certain embodiments of the subject invention, CAI prodrugs of FormulaII are provided as follows:

wherein

-   -   R₁ is selected from hydrogen, alkyl, aryl, arylalkyl,        alkylamino, dialkylamino, morpholino, piperazinyl,        alkylpiperazinyl, hydroxy, alkoxy, hydroxyalkyl, and alkoxyalkyl        or is a group of formula

-   -    wherein X is O, NH or S; Y is selected from hydrogen, alkyl,        alkylamino, dialkylamino, and hydroxyalkyl; R′ is selected from        hydrogen and a naturally occurring α-amino acid side chain; R″        is selected from hydrogen, alkyl, alkylamino, and alkylhydroxy.    -   R₂ is selected is selected from hydrogen, alkyl, aryl,        arylalkyl, alkylamino, dialkylamino, morpholino, piperazinyl,        alkylpiperazinyl, hydroxy, alkoxy, hydroxyalkyl, alkoxyalkyl,        alternatively when taken together may form a carbocyclic,        heterocyclic and heteroaryl ring

In certain other embodiments of the subject invention, CAI prodrugs ofFormula III are provided as follows:

wherein

-   -   R₃ is selected from hydrogen, alkyl, aryl, alkylaryl and        heteroaryl or is a group of formula

-   -    wherein X, Y and Z are independently hydrogen, hydroxy, alkoxy,        amino, alkylamino, dialkylamino, halogen, nitro or cyano.    -   R₄ is hydrogen, alkyl, aryl, alkylaryl or heteroaryl, or a group        of formula

-   -    wherein X, Y and Z are independently hydrogen, hydroxy, alkoxy,        amino, alkylamino, dialkylamino, halogen, nitro or cyano.

Suitable embodiments of CAI prodrugs of Formula II are those thatprovide for a high therapeutic margin of safety via conversion to CAI ata rate that yields localized therapeutic effects in target tissues (suchas ocular tissues, where esterase conversion of an esterified prodrug ofCAI is an embodiment) and compartment of administration. The preferredprodrug compounds of the subject invention are particularly advantageousbecause they provide an acceptable safety profile consistent with acuteand/or chronic ocular disease.

In one embodiment, CAI prodrugs can readily penetrate the human scleraltissue and effectively deliver the biologically active parent CAItherapeutic entity to the ocular disease target tissue compartment(s) intherapeutic concentration without causing ocular toxicity that has beenpreviously described for the CAI parent drug when administeredsystemically to cancer patients. CAI prodrugs of the present inventionmay be converted to CAI parent drug, after administration, by bothpassive (e.g. by pH dependent aqueous hydrolysis) or biologicallymediated (e.g. enzymatic) mechanisms.

In certain embodiments, CAI prodrugs of the invention can be convertedin vivo into biologically active CAI with passive hydrolytic mechanisms.For example, a CAI prodrug of Formula II can be converted to CAI in vivoby spontaneous hydrolysis. The rate of spontaneous hydrolysis of CAIprodrugs of Formula II may be tuned by appropriate choice of the R₁ andR₂ groups. Specifically, to favor the spontaneous hydrolysis ofcompounds of Formula II, one embodiment provides for compounds ofFormula II that contain nucleophilic catalytic groups, such as amino andhydroxyl functions, incorporated into either or both of the R₁ and R₂groups such that internal “anchimeric assistance” of the hydrolyticreaction is catalyzed via formation of a cyclic intermediate with a ringsize ranging from 3-7 atoms.

Alternatively, endogenous hydrolytic enzymes such as proteases andesterases in the eye may mediate the conversion of CAI amide prodrugs ofFormula II to CAI in vivo. Such hydrolytic enzymes particularly matrixmetalloproteinase are known to be upregulated in target neovasculardisease tissues for the neovascular proliferative disease indications ofrelevance specified herein. CAI amide prodrugs of Formula II may containone or more asymmetric carbon atoms and may exist in a givendiastereomeric, racemic, or enantiomeric form.

Imine and amide derivatives of amino functions and are well establishedprodrugs in the medicinal chemistry art. As described above, CAI is aknown organic compound designated as5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamideand having a structure illustrated in Formula I. The amorphous CAI freebase, which is the starting material for crystalline forms, salt forms,and other formulations that are the subject of this invention, isreadily synthesized using previously described methods (see, forexample, U.S. Pat. Nos. 4,590,201 and 5,602,156, the contents of whichare hereby incorporated by reference in their entirety) by a person whois skilled in the art of organic synthesis. CAI amide prodrugs ofFormula II are readily synthesized by persons skilled in the art forexample by reacting CAI with a reactive carboxylic acid derivative (e.g.carboxylic acid chloride or anhydride) in the presence of a base such asan organic base in an organic solvent to promote amide bond formation.Examples of carboxylic acid chlorides and anhydrides include but are notlimited to acetyl chloride, benzoylchloride, propionic anhydride,pivaloyl chloride; examples of organic bases include pyridine,4-dimethylaminopyridine and triethylamine and appropriate organicsolvents include dimethylformamide, tetrahydrofuran and dichloromethane.Compounds of Formula III are readily synthesized by persons skilled inthe art by reacting CAI with the specific ketone or aldehyde compound inan organic solvent in the presence of a catalyst or drying agent to formthe CAI ketamine or aldimine prodrugs. Examples of ketones and aldehydesinclude but are not limited to acetaldehyde, proprionaldehyde,benzaldehyde, acetophenone, benzophenone, acetone and ethylmethylketone;organic solvents include dimethylformamide, tetrahydrofuran anddichloromethane; catalysts and drying agents are exemplified bymagnesium sulfate and titanium tetraisopropoxide.

III. CAI SALTS OF THE INVENTION

Further embodiments of the present invention are novel acid additionsalt forms of CAI free base and CAI prodrugs that are used to prepare apharmaceutical formulation using any of the formulation techniques knownto the skilled artisan or as set forth herein. Particularly preferredare acid addition salt forms that, upon local administration, serve tomaintain a therapeutic, free concentration of CAI that is effective atalleviating disease pathology and have a safety profile consistent withpractical therapeutic application.

Certain embodiments of the subject invention provide acid addition saltforms of CAI derived from organic acids that have membrane stabilizingor anti-inflammatory effects including, but not limited to, cholicacids, long chain unsaturated fatty acids, and amphilic long chain andsterol derived sulfonic acids. Such salts and complexes may be preparedin a conventional manner and can exhibit superior activity, andcontrolled release profile in vivo relative to the simple CAI parentfree base. Such salts refer to non-toxic or biocompatible salts of CAIor CAI prodrugs of the invention that are generally prepared by oneskilled in the art by reacting the CAI in its free base parent form witha suitable organic or inorganic acid.

Representative organic or inorganic acids for the preparation of CAI andCAI prodrug salts of the subject invention include, but are not limitedto, the following: hydrogen borate, hydrogen bromide, hydrogen chloride,nitric acid and hydrogen nitrate, phosphoric acid or trihydrogen ordihydrogen or monohydrogen phosphate, dihydrogenphosphate, sulfuric acidor dihydrogen sulfate, and monohydrogensulfate. Representativehydrophilic organic acid salts include, but are not limited to, thefollowing: citrate, fumarate, gluconate, glutamate, lactate, maleate,mandelate, mesylate, oxalate, succinate tartrate, and valerate.Representative hydrophobic organic acid salts include, but are notlimited to, the following: benzenesulfonate, benzoate, cholate,hydroxynaphthoate, laurate, napsylate, oleate, palmoate, palmitate,salicylate, stearate, tosylate, and taurocholate. Additional salts ofessential amino acids are useful.

Particularly preferred are acid addition salt forms of CAI that, uponlocal administration, serve to maintain a therapeutic concentration offree form CAI that is effective at alleviating disease pathology withouta toxic or tissue-damaging or tissue function-impairing side effectprofile that can prohibit practical therapeutic use. Even more preferredembodiments are acid addition salt forms of CAI derived from orcomprising organic acids that have membrane stabilizing oranti-inflammatory effects such as cholic acids, long chain unsaturatedfatty acids, and amphilic long chain and sterol derived sulfonic acids.

IV. CAI COMPOUND FORMULATIONS

In a further embodiment, formulations of CAI compounds are provided fortreatment of the non-life threatening diseases, which formulations andmethods of treatment comprise the subject of this invention. Suchformulations serve to overcome bioavailability limitations of CAI as apoorly water-soluble compound and/or provide for the increased, withrespect to an aqueous solution of free base CAI, bioavailability of freeCAI drug to the targeted disease tissue and providing exposure todiseased tissue at a desired rate to achieve and maintain atherapeutically effective amount of CAI in a pharmacologically activeconcentration.

In one embodiment, aqueous CAI compound formulations administered asdescribed herein provide for immediate and complete release of CAI froma formulation. Alternatively, aqueous CAI compound formulations of theinvention, when administered, provide for a slow rate of CAI releasesuch as a controlled release or a zero order release or release of about1% to 5% of the amount of administered drug over a time period of fromabout 1 hour to about 1 month. In related embodiments, formulations ofCAI compounds of the invention include, but are not limited to,microcrystalline forms, micron and submicron solid particulateformulations, emulsion and microemulsion formulations and polymerencapsulation formulations where the drug can be dissolved or suspendedin a matrix comprising a biocompatible and/or biodegradable polymer.

Accordingly, in one embodiment,5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamideis dissolved, suspended, encapsulated or otherwise incorporated in apolymeric matrix to create a sustained-release system. The polymericmatrix may be a polylactic-coglycolic acid such as e.g. poly(D,L-lactide-co-glycolide) PLGA. In one embodiment, the matrix may bethe NOVADUR® solid polymer sustained-release drug delivery system. Thesystem and/or matrix may be free of a preservative. The system may alsocontain other components described below.

In another embodiment, the CAI compound formulations of the inventionprovide a slow rate of CAI release in the target disease tissue over a1-week to 3-year period after a single administration. Certain CAIcompound formulations of the invention provide pharmacologicallyeffective concentrations of CAI in the target disease tissue over a 15minute to one-week period week after single administration.

The CAI compound formulations described below can be prepared inproportions of ingredients (CAI compound and pharmaceutically acceptableingredients such as excipients, surface-active agents, solvents, and thelike) of which are determined by the solubility and chemical nature ofthe CAI compound or formulation, chosen route of administration, andstandard medical practice. For example, microparticle formulations ofCAI can be further prepared in an aqueous formulation, for example inthe presence of a pharmaceutically acceptable surfactant or apharmaceutically acceptable surface-active agent.

A. Emulsion Formulations

Emulsion formulations of CAI compounds of the invention include, but arenot limited to, compositions prepared using formulation processescomprising a lipid, surfactant, and solvent that are readily performedby one skilled in the art of drug formulation. In a one embodimentemulsion formulations of CAI compounds to be administered as describedherein, provide for immediate or short term release of at least 90% ofthe amount of CAI in the formulation and providing pharmacologicallyeffective concentrations of CAI in the target disease tissue over a 15minute to one-week period week after single administration.

In general emulsion formulations of CAI compounds of the invention areprepared using a selection of the following five classes of excipients(Pouton, C. W. Eur. J. Pharm. Sci. 11 Suppl 2: S93-98, 2000): 1) puretriglyceride oils; 2) mixed glycerides; 3) lipophilic surfactants; 4)hydrophilic surfactants; and 5) water-soluble cosolvents. Examples oflipid based drug formulations applicable to CAI, CAI prodrug and saltsthereof include emulsions prepared for the lipophilic immunosuppressivedrug cyclosporine (Fahr, A. et al. Clin. Pharmacokinet. 24, 472-495,1993; Fricker, G. et al. Br. J. Pharmacol. 117, 217-223, 1996), andhalofantrine a lipophilic small molecule antimalaria drug (Porter, C.J., et al. J. Pharm. Sci. 85, 357-361, 1996).

In certain embodiments, emulsion formulations of the inventioncomprising liquid droplets of formulation comprising CAI or a CAIcompound, which droplets are surface stabilized dispersions of dropletsof sizes less than 10 micrometers in aqueous medium. Alternatively, thesubject invention's emulsion formulations can be locally delivered to atarget tissue as an oil-in-water microemulsion of droplet sizes lessthan 100 nanometers. These microemulsions are composed of CAI, CAIprodrugs and salts thereof dissolved in a mixture of lipophilic vehicleand admixed with surfactants and cosurfactants.

In other embodiments, both these end results (lipophilic carrier andadmixture with surfactants and cosurfactants) are achieved from apreconcentrated or a concentrated solution of a CAI compound in avehicle that forms oil-in-water emulsion of droplet sizes less than 10micrometers spontaneously upon mixing with bodily fluids (i.e., lacrimalfluid, vitreal fluid, scleral fluid, blood, cerebral spinal fluid,vaginal fluid, mucous). Alternatively, emulsion formulations of theinvention can be achieved from a preconcentrated or a concentratedsolution of a CAI compound in a vehicle that forms oil-in-watermicroemulsion of droplet sizes less than 100 nanometers spontaneouslyupon mixing with bodily fluids. In this way, in-situ generation of thesmall size droplets is facilitated wherein application of energy is notrequired.

Emulsion formulations of CAI compounds are generally selected andprepared as described below. First, suitable pharmaceutically acceptableoil or triglyceride vehicles preferably a readily availablepharmaceutically acceptable oil or pharmaceutically acceptabletriglyceride vehicle (such as pure triglyceride oils) is selected from acommercial source e.g. soybean oil (Fluka, Buchs, Switzerland), sesameoil (Fluka, Buchs, Switzerland), Miglyol® (810, 812, 818, Hulls,Puteaux, France) and other candidates selection of which is preferablybased on respectively highest solubility and longest term of stabilityof the pure CAI, CAI salts or prodrugs as active ingredient in the oil,solubility and stability determined in preformulation evaluation bydissolving CAI in the vehicle, +/−surfactant, +/−water (optionallycontaining a buffer salt) to determine saturation concentrations of CAIas a function of temperature and then determining the stability of CAIas a function of time (1 day, 2 days, 4 days, 1 week 2 weeks, 4 weeks, 1month, 2 months, and monthly thereafter, each at constant temperaturesof about 20° C., 30° C., and 40° C. Formulations containing hydrophobicsurfactants and hydrophilic surfactants are then prepared with an oilvehicle composition range of 25-45%, hydrophobic surfactant(s) range of25-45% and hydrophilic co-surfactant(s) in a range of 10-20%.

Contemplated hydrophobic surfactants for use in the preparation ofemulsion formulations of the invention include, but are not limited to,glyceryl monostearate and Pluronic® (e.g., Pluronic®F68, Pluronic®F127,Sigma, St. Louis Mo.). Contemplated hydrophilic co-surfactants includebut are not limited to, Tween® (e.g., Tween® 20, Tween® 40, Tween® 80,Sigma, St. Louis Mo.) and Labrasol® (Gattefosse, Saint Priest, France).Preferred emulsion formulations of the invention exhibit small submicrondroplet size, high CAI active ingredient stability, and high CAI activeingredient load.

Alternatively, in accordance with the subject invention, a CAI compoundcan be formulated as an aqueous suspension of surface stabilizedparticles. The particle size distribution of such suspensionformulations is carefully maintained to less than 10 micrometers so thatlocal irritation to the target tissue (for example, tissue of the eye)is avoided. For example, one embodiment of the invention provides apreparation of concentrated solution of a CAI compound. Specifically,the CAI compound is dissolved or suspended directly in vehicles thatform a suspension of surface stabilized CAI compound with particles ofsizes less than 10 micrometers. The vehicles provide therapeuticallyeffective concentrations of pharmacologically active CAI at the targetdisease site upon local administration to the disease target tissue ortissue compartment by administration methods described herein. Surfacestabilization of the particles in these embodiments is provided by apharmaceutically acceptable surface active agent, for example a surfaceactive agent selected from the group consisting of lecithin, charged oruncharged phospholipids, polymeric surfactants, non-polymericsurfactants, charged surfactants (e.g., anionic surfactants or cationicsurfactants), uncharged surfactants (e.g., polyether surfactants such aspolyethyleneglycol-containing surfactants andpolyethyleneglycol-co-polypropyleneglycol-containing surfactants), oneor more of bile acids and their salts, or combinations thereof.

B. Beta-Cyclodextrin Complexes

Alternatively, the CAI compounds of the invention, such as e.g. CIA, canbe prepared as a molecular complex with beta-cyclodextrin or itsderivatives. Furthermore, an alternative formulation consists ofmolecular complex of a CAI compound with one or more pharmaceuticallyacceptable lipids selected from a group of lecithin, charged oruncharged phospholipids, polymeric surfactants, non-polymericsurfactants, charged surfactants, uncharged surfactants, one or more ofbile acids and their salts, triglycerides, or other lipophilic solventsor combinations thereof.

Thus, in certain embodiments, CAI may be in a molecular complexformulation with 1 to 60 wt % hydroxypropyl β-cyclodextrin informulation, wherein the formulation further contains at least acellulose derivative, and a surfactant. In other embodiments, theformulation contains glutamate and/or L-phenylalanine. The cellulosederivate may be hydroxypropyl methyl cellulose. These complexes may havea diameter of less than about 10 micrometers.

C. Microparticle Formulations

“Microparticlulate forms of a CAI compound” refers to solid particulateforms of CAI free base, CAI prodrugs and salts thereof with particlesizes of 10 micrometers or less. Further, “microparticlulate forms of aCAI compound” refers to respective amorphous or crystalline forms CAI,CAI prodrugs and salts thereof as well as mixtures of CAI, CAI prodrugsand salts thereof with other additives (e.g. polymers, surfactants) thatyield particulate forms of 10 micrometers or less.

Microparticle formulations of CAI compounds of the invention include,but are not limited to, compositions prepared using controlledprecipitation formulation processes readily selected and implemented byone skilled in the art of drug formulation. For example, by dissolving aCAI compound in a solvent such as acetone or ethanol or tetrahydrofuranor DMSO to form a solution and then adding said solution to anon-solvent such as water in which the solvent has solubility, or addingsuch non-solvent slowly with stirring to the solution, each methodoptionally in the presence of a pharmaceutically acceptable surfaceactive agent or combination of agents in either the solvent ornon-solvent or in both, followed by removal of the solvent, such as byevaporation or diafiltration to provide a suspension of microparticlesin an aqueous medium. The particles can be isolated by lyophilization ofthe suspension, for example in vials optionally in the presence of anoxygen-free gas such as nitrogen or argon after removal of the water,optionally after addition of a carbohydrate such as lactose or mannitolwhich can form a solid matrix in which the solid particles aredispersed. The particles can be resuspended into an aqueous medium uponaddition of such aqueous medium. In an embodiment, microparticulateformulations of a CAI compound to be administered as described herein,provide for immediate release of CAI providing pharmacologicallyeffective concentrations of CAI in the target disease tissue over aone-week to three-year period after a single administration.

In one embodiment, microparticles of CAI of the invention includepolymeric microspheres that encapsulate CAI, CAI prodrugs and saltsthereof of the invention. Microparticle formulations of CAI include, butare not limited to, compositions prepared using controlled precipitationformulation processes readily selected and implemented by one skilled inthe art of drug formulation. A controlled microparticle precipitationprocess is exemplified as follows.

CAI free base or a respective CAI acid addition salt form or prodrug isdissolved in a suitable organic solvent (e.g. ethanol,N-methylpyrrolidine) and then mixed with stabilizers, as exemplified bySPG (0.218 sucrose; buffer salts comprising 0.0038 M monobasic, 0.0072 Mdibasic potassium phosphate; 0.0049 M potassium glutamate). Polymer(e.g. polyvinylpyrrolidone) and sugar (e.g. lactose) matrix-formingcomponents are optionally added. Micron and submicron microparticles ofCAI formulation are then prepared by controlled precipitation at anoptimized temperature under low frequency sonication followed.Sonication of the contents of the vessel containing the components ofsuch formulation can also be accomplished using ultrasound frequencies,for example by placing the vessel containing the formulation in anultrasound bath and applying ultrasound energy. The CAI microparticleformulation(s) are collected, for example by centrifugation such as at10,000 g and characterized by digital optical light microscopy and laserlight scattering and Scanning Electron Microscopy (SEM) for particlesizing. Contents of the formulation are analyzed, for example by usingHPLC for content and stability of such content, and by using USPdissolution techniques to measure the release rate of the CAI drug invitro. Thus, in one embodiment, CAI is provided as a sonicatedmicroparticle. In certain embodiments, especially when used to treatuveitis, the sonicated CAI microparticles may be of a size less thanabout 10 micrometers.

In yet another embodiment, the CAI microparticle formulation is preparedusing methods that are standard in the art, such as by entrapment orencapsulation of CAI free base, CAI salt form or prodrug drug moleculeswithin a matrix of polymeric microspheres of sizes less than 50micrometers, wherein the polymeric matrix material is selected from thegroup of albumin, polylactic-coglycolic acid, polymethacrylic acids andsalts, polyethyleneglycol, natural polymeric materials, syntheticpolymeric materials, charged polymeric materials, and unchargedpolymeric materials, and combinations thereof.

Suitable polymer encapsulation systems include polylactic-coglycolicacid (PLGA) microsphere formulations as exemplified by the formulationof octreotide in Sandostatin-LAR® (See Sandostatin LAR® prescribinginformation) which is administered as a once-a-month depot injectabletherapeutic.

D. Liposomal Formulations

In addition, the compounds of the present invention can also beadministered in the form of liposomes or the like. Disintegratorsinclude, without limitation, delivery systems such as small unilamellarvesicles, large unilamellar vesicles and multilamellar vesicles.Liposomes can be formed from a variety of phospholipids, such aslecithin and phosphatidylcholines, optionally with added cholesteroland/or stearylamine.

D. Pharmaceutical Formulations, in General

CAI compounds of the invention may be provided in formulations thatcontain pharmaceutically acceptable auxiliary substances as required toapproximate physiological conditions, such as pH adjusting for exampleto adjust pH of an aqueous medium, for example to about pH 7.4 andbuffering agents to maintain such pH, tonicity adjusting agents whichcan be useful, for example, to adjust tonicity of a formulation tophysiological ionic strength, wetting agents and the like, for example,sodium acetate, sodium lactate, sodium chloride, potassium chloride,calcium chloride, sorbitan monolaurate, triethanolamine oleate, etc.

The concentration of the CAI compounds of the invention in theirrespective pharmaceutical formulations can vary widely (% w/v), i.e.,from less than about 0.1%, usually at or at least about 2% to as much as20% to 50% or more by weight, and will be selected primarily by desireddosage levels, fluid volumes, viscosities, etc., in accordance with theparticular mode and route of administration selected and target tissue.Preferably, the concentration of CAI compound in the aqueous,therapeutic formulations of the invention range from about 0.01 mg/mL toabout 100 mg/mL. More preferably, the concentration of CAI compound inthe aqueous, therapeutic formulations of the invention for the localizedtreatment of ocular diseases range from about 1 mg/mL to 60 mg/mL offormulation.

The pharmaceutical formulations of the invention may includedelivery-enhancing agents that can be used alone, in combination witheach other, or in combination with another delivery-enhancing agent. Theterm “delivery enhancing agents” includes agents that facilitate thetransfer CAI to the target cell. Examples of such delivery enhancingagents include, but are not limited to, fatty acid esters, surfactants,detergents, alcohols, glycols, surfactants, bile salts, heparinantagonists, cyclooxygenase inhibitors, hypertonic salt solutions,acetates.

As contemplated herein, alcohols that can act as delivery enhancingagents include, without limitation, the aliphatic alcohols such asethanol, N-propanol, isopropanol, butyl alcohol, acetyl alcohol. Glycolsthat can act as delivery enhancing agents in accordance with the subjectinvention include, but are not limited to, glycerine, propyleneglycol,polyethyleneglycol and other low molecular weight glycols such asglycerol and thioglycerol. Acetates such as acetic acid, gluconic acid,and sodium acetate are further examples of delivery-enhancing agents.Hypertonic salt solutions like 1M NaCl are also examples ofdelivery-enhancing agents.

Examples of pharmaceutically acceptable surfactants that can be providedin pharmaceutical formulations of the invention include, withoutlimitation, sodium dodecyl sulfate (SDS) and lysolecithin, polysorbate80, nonylphenoxypolyoxyethylene, lysophosphatidylcholine,polyethylene-glycol 400, polysorbate 80, polyoxyethylene ethers,polyglycol ether surfactants and DMSO. Bile salts such as taurocholate,sodium tauro-deoxycholate, deoxycholate, chenodesoxycholate, glycocholicacid, glycocheno-deoxycholic acid and other astringents like silvernitrate may be used. Heparin-antagonists like quaternary amines such asprotamine sulfate may also be used. Cyclooxygenase inhibitors such assodium salicylate, salicylic acid, and non-steroidal anti-inflammatorydrug (NSAIDS) like indomethacin, naproxen, diclofenac may be used.

Examples of pharmaceutically acceptable detergents that can be providedin pharmaceutical formulations of the invention can be selected fromanionic, cationic, zwitterionic, and nonionic detergents. Exemplarypharmaceutically acceptable detergents include but are not limited totaurocholate, deoxycholate, taurodeoxycholate, cetylpyridium,benalkonium chloride, ZWITTERGENT-3-14 detergent, CHAPS(3-{(3-Cholamidopropyl)dimethylammoniol}-1-propanesulfonate hydrate,Aldrich), Big CHAP, Deoxy Big CHAP, TRITON™ X-100 detergent, C12E8,Octyl-B-D-Glucopyranoside, PLURONIC® F68 detergent, TWEEN® 20 detergent,and TWEEN® 80 detergent.

The concentration of a delivery-enhancing agent in a pharmaceuticalformulation of the invention will depend on a number of factors known toone of ordinary skill in the art. Such factors include the particulardelivery-enhancing agent being used, the buffer, pH, target tissue ororgan and mode of administration. In certain embodiments of theinvention, the concentration of the delivery-enhancing agent will be inthe range of 1% to 50% (v/v), preferably 10% to 40% (v/v), and mostpreferably 15% to 30% (v/v). Preferably, the detergent concentration inthe final formulation administered to the patient is about 0.5 to about2 times the critical micellization concentration (CMC).

Excipients can be included in the formulations of the invention.Examples include cosolvents, surfactants, oils, humectants, emollients,preservatives, stabilizers, and antioxidants. A pharmacologicallyacceptable buffer may be used, e.g., tris or phosphate buffers.Effective amounts of diluents, additives, and excipients are thoseamounts, which are effective to obtain a pharmaceutically acceptableformulation in terms of solubility, stability, and biological activity.

Formulations of the invention may also include, depending on theformulation and mode of delivery desired, pharmaceutically-acceptable,non-toxic carriers or diluents, which include vehicles commonly used toform pharmaceutical compositions for animal or human administration. Thediluent is selected so as not to unduly affect the biological activityof the combination. Examples of such diluents that are especially usefulfor injectable formulations are water, the various commerciallyavailable pharmaceutically-acceptable saline, organic, or inorganic saltsolutions, Ringer's solution, dextrose solution, and Hank's solution. Inaddition, the pharmaceutical composition or formulation may includeadditives such as other carriers; adjuvants; or nontoxic,nontherapeutic, nonimmunogenic stabilizers and the like.

Carrier employed may be, for example, either a solid or liquid. Examplesof solid carriers include lactose, terra alba, sucrose, talc, gelatin,agar, pectin, acacia, magnesium stearate, stearic acid, and the like.Examples of liquid carriers include syrup, peanut oil, olive oil, water,and the like. Similarly, a carrier for oral use may include time delaymaterial well known in the art, such as glyceryl monostearate orglyceryl distearate alone or with a wax.

In certain embodiments, pharmaceutical formulations comprise a CAIcompound (preferably, CAI itself) and a carrier such as inclusioncompound host materials. The “inclusion compound host materials” asdescribed herein, interact with the CAI compound to increase aqueoussolubility, increase chemical stability, and/or enhance drug (such asCAI compound) delivery to and through biological membranes. It isbelieved that by providing a carrier such as inclusion compound hostmaterials, a stabilized CAI compound molecule can be safely delivered toa patient at a dosage that will not induce toxicity. In addition, suchcarrier materials can include coating materials (i.e., enteric-coatings)that allow dissolution of the coating in an alkaline environment such asin the intestines.

Inclusion compound host materials that can be used in accordance withthe subject invention include those disclosed in U.S. Published PatentApplication No. 2004/0033985, incorporated herein by reference in itsentirety. Contemplated inclusion compound host materials includeproteins (such as albumin), crown ethers, polyoxyalkylenes,polysiloxanes, zeolites, cholestyramine, colestipol, colesevelam,colestimide, sevelamer, cellulose derivatives, dextran derivatives,starch, starch derivatives, and pharmaceutically acceptable saltsthereof. Contemplated cellulose derivatives and dextran derivativesinclude DEAE-cellulose, guanidinoethylcellulose, or DEAE-Sephadex.Favorable starches or starch derivatives to be included in thecompositions of the invention include cyclodextrin, retrograded starch,degraded starch, a combination of retrograded and degraded starch,hydrophobic starch, amylase, starch-diethylaminoethylether, andstarch-2-hydroxyethyl ether.

According to the subject invention, aqueous CAI compound formulationscomprise CAI and inclusion compound host materials selected from, butnot limited to, cyclodextrin and/or its derivatives (i.e., methylβ-cyclodextrin (M-β-CD), hydroxypropyl β-cyclodextrin (HP-β-CD),hydroethyl β-cyclodextrin (HE-β-CD), polycyclodextrin, ethylβ-cyclodextrin (E-β-CD) and branched cyclodextrin). As one skilled inthe art will appreciate, any cyclodextrin or mixture of cyclodextrins,cyclodextrin polymers, or modified cyclodextrins can be utilizedpursuant to the present invention. Cyclodextrins are available fromWacker Biochem Inc., Adrian, Mich. or Cerestar USA, Hammond, Ind., aswell as other vendors. Formation of inclusion complexes usingcyclodextrin or its derivatives protects the constituent (i.e., CAIcompound) from loss of evaporation, from attack by oxygen, acids,visible and ultraviolet light and from intra- and intermolecularreactions.

The general chemical formula of cyclodextrin is (C₆O₅H₉)_(n). Thecontent of inclusion compound host materials in compositions of thesubject invention can range from about 1 to 80 wt %. Preferably, thecontent of inclusion compound host materials in compositions of theinvention range from about 1 to 60 wt %. The actual amount of theinclusion compound host materials used will depend largely upon theactual content of CAI compound and therapeutic agents, if any, used inpreparing compositions of the invention.

Preferably, aqueous pharmaceutical formulations of CAI compounds can beprepared as molecular complexes with beta-cyclodextrin or itsderivatives. Furthermore, an alternative pharmaceutical formulationconsists of molecular complex of CAI compound(s) with one or more lipidsselected from a group of lecithin, charged or uncharged phospholipids,polymeric surfactants, non-polymeric surfactants, charged surfactants,uncharged surfactants, one or more of bile acids and their salts,triglycerides, or other lipophilic solvents or combinations thereof

The CAI compound and aqueous formulations thereof of the subjectinvention can be formulated according to known methods for preparingpharmaceutically useful compositions. Formulations are described in anumber of sources, which are well known and readily available to thoseskilled in the art. For example, Remington's Pharmaceutical Science(Martin E W [1995] Easton Pa., Mack Publishing Company, 19^(th) ed.)describes formulations that can be used in connection with the subjectinvention. Formulations suitable for parenteral administration include,for example, aqueous sterile injection solutions, which may containantioxidants, buffers, bacteriostats, and solutes, which render theformulation isotonic with the blood of the intended recipient; andaqueous and nonaqueous sterile suspensions, which may include suspendingagents and thickening agents. The formulations may be presented inunit-dose or multi-dose containers, for example sealed ampoules andvials, and may be stored in a freeze dried (lyophilized) conditionrequiring only the condition of the sterile liquid carrier, for example,water for injections, prior to use. Extemporaneous injection solutionsand suspensions may be prepared from sterile powder, granules, tablets,etc. It should be understood that in addition to the ingredientsparticularly mentioned above, the formulations of the subject inventioncan include other agents conventional in the art having regard to thetype of formulation in question.

Certain embodiments of the invention may be formulated for localinstillation in or around tissue or organs, where for specific targetedlocal therapy for non-ocular body sites, parenteral therapy is used toachieve safe and effective local drug concentrations into a definedtarget lesion or disease area, or tissue(s), or organ(s), or bodycompartment(s), wherein routes of administration of formulations of CAIcompounds can include transcutaneous, subcutaneous, intradermal,intrathecal, intracerebellar, intramuscular, intra-articular orintravenous, when such direct application is practical and clinicallyindicated.

Alternatively, certain embodiments of the invention may be formulatedfor systemic administration to a patient diagnosed with a non-lifethreatening disease.

The CAI compounds of the invention can be administered in topicalformulations or polymer matrices, hydrogel matrices, polymer implants,or encapsulated formulations to allow slow or sustained-release of thecompositions. A particularly suitable formulation is a suspension orsolution of the delivery system in a topical ocular formulation, such aseye drops.

Topical applications may be formulated in carriers such as hydrophobicor hydrophilic bases to form ointments, creams, lotions, in aqueous,oleaginous or alcoholic liquids to form paints or in dry diluents toform powders. Such topical formulations can be used to treat oculardiseases as well as inflammatory diseases such as rheumatoid arthritis,psoriasis, contact dermatitis, delayed hypersensitivity reactions, andthe like.

Certain embodiments of the invention are designed to locally deliverbioactive CAI into target eye tissue from implants to be injected in oraround the eye, for sustained delivery of bioactive CAI to the eye.Alternatively, delivery vehicles are designed in the form of ointments,gels, viscous suspensions, or viscous emulsions that releases the drugupon administration into or around eye for prolonged periods to elicitthe desired pharmacological action. The dosage form can also be a freeflowing sterile suspension for topical administration or free flowingsterile suspensions for injection into or around eye.

When the CAI compound is formulated as a solution or suspension, thedelivery system is in a pharmaceutically acceptable carrier, preferablya pharmaceutically acceptable aqueous carrier, suitable for local tissuecompatibility. A variety of aqueous carriers may be used, e.g., water,buffered water, 0.8% saline, 0.3% glycine, hyaluronic acid and the like.Such delivery systems may be sterilized by conventional, well knownsterilization techniques e.g., by heat or steam sterilization offormulations in sealed vials at about 130° C. for at least 15 minutes,or liquid formulations may be sterile filtered, e.g., by filtrationthrough a 0.2 micron filter in a sterile pharmaceutic environment.

V. ROUTES OF ADMINISTRATION

The CAI compounds of the invention, and formulations thereof, can beused in a variety of routes of administration, including, for example,orally-administrable forms such as tablets targeting a specific localtissue e.g. gastrointestinal tissues, capsules or the like, or viainjectable (e.g., parenteral, ocular, intravenous, subcutaneous,intramuscular), topical (e.g., dermal, ocular, transdermal, nasal),buccal, pulmonary (e.g., inhalation), suppository, or other route. Inone embodiment, they can be in unit dosage form, namely, in physicallydiscrete units suitable as unitary dosages for human consumption, eachunit containing a predetermined quantity of active ingredient calculatedto produce the desired therapeutic effect in association with one ormore pharmaceutically acceptable other ingredients, i.e., diluent orcarrier.

The routes of localized administration for the treatment of oculardiseases using the CAI compound or formulations thereof, of theinvention include ophthalmic artery administration, subretinalinjection, intravitreal injection, and periocular injection orjuxtascleral administration. In the case of angiogenic diseases such asage-related macular degeneration and diabetic retinopathy, theformulations of the present invention are administered over a course oftreatment ranging from weeks to years. The routes of administration forthe treatment of such diseases include ophthalmic artery administration,subretinal injection, intravitreal injection, and periocular injectionor juxtascleral administration. Sustained-release formulations such asimplants would also be appropriate for the treatment of such long-termdisease indications. These formulations may also be administered incombination with other anti-angiogenic agents.

In one embodiment of the invention, a CAI compound can be injectedintraocularly using intravitreal (into the vitreous), subconjuctival(into the subconjuctival), subretinal (under the retina), or retrobulbar(behind the eyeball) injection. For subconjuctival injection, a CAI dosein the range of about 0.1 ng/ml to about 10 mg/ml may be used. Forintravitreal injection, a CAI dose in the range of about 0.1 ng/0.1 mlto about 10 mg/0.1 ml may be used. For retrobulbar injection, a CAI dosein the range of about 1 ng/ml to about 10 mg/ml may be used. Forsubretinal injection, a CAI dose in the range of about 0.1 ng/0.1 ml toabout 10 mg/0.1 ml may be used.

A. Time-Released Delivery Route

Slow or extended-release delivery systems, such as delivery systemscomprising a biopolymer (biological-based systems), liposomes, colloids,resins, and other pharmaceutical acceptable polymeric delivery systemsor compartmentalized reservoirs, can be utilized with the compositionsdescribed herein to provide a continuously releasing or long termreleasing source of therapeutic compound. Such slow release systems areapplicable to formulations for delivery via topical, intraocular, oral,and parenteral routes. Delivery to areas within the eye, in situ can beaccomplished by injection such as by using a needle and syringe, by useof a cannula or other invasive device designed to introduce preciselymetered amounts of a desired formulation to a particular compartment ortissue within the eye (e.g. anterior or posterior chamber, uvea orretina). A solid, semisolid, or liquid implant can be deliveredsubretinally using the instrumentation and methods described in U.S.Pat. Nos. 5,817,075 and 5,868,728, the contents of which are herebyincorporated by reference in their entirety.

Sustained-Release Systems Particularly Useful for the Treatment ofUveitis.

In certain embodiments of the invention, the CAI compound is formulatedto be part of a sustained-release system. Such sustained release systemsmay be particularly useful for the treatment of uveitis, especiallyposterior infectious uveitis. Thus, in one embodiment,5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamidemay be formulated to be part of a sustained-release system. In anotherembodiment, the CAI compounds for Formula II or III may be formulated tobe part of sustained-release system.

The sustained-release systems may be formulated to contain one or moreof anti-inflammatory agent, an antibacterial agent, or a steroid (suchas e.g. a glucocorticoid). As such, the sustained release systems may besuitable for use in combination therapy (see Combinatorial Therapiesbelow). Thus, in one embodiment of the invention CAI alone or incombination with one or more of anti-inflammatory agent, anantibacterial agent, or a steroid in a sustained release system may beused to treat posterior infectious uveitis such as e.g. posteriorinfectious uveitis caused by toxoplasmosis, toxocara and visceral larvamigrans.

The sustained-release system may be an implant. The implant may be asustained release device or a sustained release-system that dissolvesover time. In one embodiment, the sustained-release drug delivery systemincludes a polymer. In one embodiment, the polymer may be configured sothat it dissolves over time (such as e.g. for up to 6 months) whenimplanted into the eye. In certain embodiments, the polymer may comprisea polylactic-coglycolic acid (PLGA). In a specific embodiment, thepolymer may comprise poly (D,L-lactide-co-glycolide) PLGA. In oneembodiment, the drug delivery system may also comprise a glucocorticoid.In one embodiment, the drug delivery system includes dexamethasone. Theamount of dexamethasone in the system may vary. In certain embodiments,the amount of dexamethasone may range from about 0.1 to about 0.9 mg,alternatively from about 0.5 to about 0.8 mg, alternatively about 0.7mg. In one embodiment, the CAI compound (e.g.5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide)is formulated to be part of an OZURDEX® (dexamethasone) intravitrealimplant. Exemplary polymer sustained-release drug delivery systemcontaining dexamethasone into which the CAI compound may be incorporatedare disclosed in U.S. Pat. Nos. 6,726,918; 6,899,717; 7,033,605;7,767,223; 8,034,366; 8,034,370; 8,043,628; 8,063,031; 8,088,407 and8,506,987, the disclosures of which are incorporated in their entiretyas they pertain to sustained-release drug delivery systems. In certainembodiments, the dexamethasone and CAI compound may be incorporated intoa poly (D,L-lactide-co-glycolide) PLGA intravitreal polymer matrix. Thismatrix may be free of a preservative. In certain embodiments, the matrixmay be the NOVADUR® solid polymer sustained-release drug deliverysystem.

The polymer sustained-release drug delivery system may have a variety ofdifferent configurations. In certain embodiments, the implant is rodshaped.

In one embodiment, the sustained-release drug delivery system is adevice configured for implantation into the eye. Such devices may have avariety of shapes. In certain embodiments, the devices are injectable,non-erodible, intravitreal implants. Suitable sustained-release deviceare configured such that a pharmaceutically acceptable amount of CAI isreleased daily. In addition to CAI, these devices may also contain otherpharmaceutically active compounds such as, for example, one or more ofanti-inflammatory agent, an antibacterial agent, or a steroid (such ase.g. a glucocorticoid). In one embodiment, the device contains CAI andfluocinolone acetonide. Exemplary suitable sustained release devices aredisclosed, for example, in U.S. Pat. Nos. 5,378,475; 5,773,019;6,217,895; 6,375,972; 6,548,078; 8,252,307; 8,574,659 and 8,871,241, thedisclosures of which are incorporated in their entirety as they pertainto sustained-release drug delivery devices. In one embodiment, the CAIcompound (e.g.5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide)is formulated to be part of an ILUVIEN® (fluocinolone acetonide)intravitreal implant.

When used as part of a sustained-release drug delivery system, the CAIcompound may be provided as a sonicated microparticle or in a molecularcomplex. In one embodiment, the CAI compound is provided as molecularcomplex in formulation with 1 to 60 wt % hydroxypropyl β-cyclodextrinand one or more of glutamate, L-phenylalanine, a cellulose derivative, asurfactant or combinations thereof. The cellulose derivate may behydroxypropyl methyl cellulose.

In certain embodiments of the invention, the CAI compound is formulatedto be part of a sustained-release system for the treatment uveitis, e.g.posterious infectious uveitis. When used in such a formulation, theamount of the CAI compound may range from about 100 to about 200 mcg,alternatively from about 150 to about 220 mcg, alternatively from about150 to 200 mcg, alternatively about 150 mcg, or alternatively about 200mcg.

In a time-release formulation, the microsphere, capsule, liposome, etc.may contain a concentration of CAI that could be toxic if it wereadministered as a bolus dose. The time-release administration, however,is formulated so that the concentration released over any period of timedoes not exceed a toxic amount. This is accomplished, for example,through various formulations of the vehicle (coated or uncoatedmicrosphere, coated or uncoated capsule, lipid or polymer components,unilamellar or multilamellar structure, and combinations of the above,etc.). Other variables may include the patient'spharmacokinetic-pharmacodynamic parameters (e.g., body mass, gender,plasma clearance rate, hepatic function, genetic makeup, etc.).Depending upon the amount of CAI provided in the formulation and therelease rate of the CAI, a patient could be dosed with CAI over a periodof years from a single implant or injection comprising a CAI compound.As illustrative but non-limiting examples, a capsule or a device, e.g.,for implantation use, can be loaded with a CAI-based formulationcomprised of a concentration within the range of 1 to 200 mg of activeCAI ingredient (e.g., 1 mg, 2 mg, 5 mg, 10 mg, 40 mg, 80 mg, 100 mg, 121mg, 155 mg, 200 mg); if the capsule is formulated to release a few(e.g., 1 to 100) micrograms of CAI active ingredient drug per day, thepatient's disease could be treated for a period ranging from about7-1000 days (1-week to 3-years) via a single administration of thecapsule or device. Such a formulation provides benefits which includeaccurate dosing with heightened patient convenience, because frequentintervention or administration is not required, rather administrationcan be done in some cases only once or twice a decade or even lessfrequently. The formation and loading of microspheres, microcapsules,liposomes, etc. and their ocular implantation are standard techniquesknown by one skilled in the art, for example, the use a ganciclovirsustained-release implant to treat cytomegalovirus retinitis, disclosedin Vitreoretinal Surgical Techniques, Peyman et al., Eds. (MartinDunitz. London 2001, chapter 45); Handbook of Pharmaceutical ControlledRelease Technology, Wise, Ed. (Marcel Dekker, New York 2000), therelevant sections of which are incorporated by reference herein in theirentirety.

B. Dosage Forms

Depending on the clinical needs of a patient, formulations of thesubject invention (see Section IV above) are prepared to either locallydeliver the bioactive CAI immediately after administration or forbioactive CAI release into a target tissue (for example, eye tissue)from its vehicle in a sustained manner over a period of time suitablefor the desired pharmacological action.

An effective quantity of the compound of interest is employed intreatment. The dosage of compounds used in accordance with the inventionvaries depending on the compound and the condition being treated. Forexample, the age, weight, and clinical condition of the recipientpatient, and the experience and judgment of the clinician orpractitioner administering the therapy are among the factors affectingthe selected dosage. Other factors include the route of administration,the patient, the patient's medical history, the severity of the diseaseprocess, and the potency of the particular compound. The dose should besufficient to ameliorate symptoms or signs of the disease treatedwithout producing unacceptable toxicity to the patient. In general, aneffective amount of the compound is that which provides eithersubjective relief of symptoms or an objectively identifiable improvementas noted by the clinician or other qualified observer. In the case ofangiogenic diseases such as age-related macular degeneration anddiabetic retinopathy, the formulations of the present invention areadministered over a course of treatment ranging from weeks to years.

C. Combinatorial Therapies

A further embodiment of the subject invention provides for the localadministration of a CAI compound concurrently with other pharmacologicaltherapies for the treatment of non-life threatening diseases. Forexample, a CAI compound of the invention can be administeredconcurrently with other clinical therapies such as the concurrentadministration with anti-angiogenic compounds (e.g., combretastatin,angiostatin, endostatin, vitaxin, 2ME₂, anecortave, squalamine,Macugen®, Lucentis®, PEDF), which may have diverse mechanisms of action(e.g. VEGF neutralization, tyrosine receptor kinase inhibition),glucocorticoids and non-steroidal anti-inflammatory drugs.

The present invention provides for the concurrent, localizedadministration of therapies and a CAI compound or formulations thereof,wherein the therapy addresses a non-life threatening disease (such ascertain non-cancerous proliferative and angiogenic diseases) other thanthat treated by the CAI compound or formulations thereof, of theinvention. For example, one embodiment of the invention provides thetopical delivery of a CAI prodrug and novel formulations for treatingsevere dermatological diseases including severe psoriasis, excema androsacea and local intrarticular administration for severe arthritis byinhibiting vascular and inflammatory cell proliferation.

A further embodiment of the subject invention provides for the treatmentof neovascular and edematous ocular diseases by ocular administration ofa CAI compound, as described herein, in combination with otherpharmacological anti-angiogenesis therapies. Contemplatedanti-angiogensis therapies comprise those which include, but are notlimited to, glucocorticoids (such as, dexamethasone, fluoromethalone,administration of medrysone, budesonide, betamethasone, fluocinalone,triamcinolone, triamcinolone acetonide, prednisone, prednisolone,hydrocortisone, rimexolone, and pharmaceutically acceptable saltsthereof), anecortave acetate, VEGF-binding molecules (oligonucleotideaptamers (e.g. Macugen®), protein antibodies (e.g. Lucentis®), tyrosinereceptor kinase inhibitors including but not limited to vascularendothelial growth factor (VEGF), platelet derived growth factor (PDGF)and fibrobrast growth factor (FGF) receptors, other direct or indirectgrowth factor inhibitors including somatostatin receptor agonists(inhibiting release of Growth Hormone and IGF-1), RNAi oligonucleotidetranscription inhibitors of ocular disease molecular targets includinggrowth factors described above.

Active agents suitable for concurrent administration with a CAI compoundor formulation thereof include, but are not limited to, anti-infectives,including, without limitation, antibiotics, antivirals, and antifungals;antiallergenics and mast cell stabilizers; steroidal and non-steroidalanti-inflammatory agents; cyclooxygenase inhibitors, including, withoutlimitation, Cox I and Cox II inhibitors; combinations of anti-infectiveand anti-inflammatory agents; anti-glaucoma agents, including, withoutlimitation, adrenergics, beta-adrenergic blocking agents, α-adrenergicagonists, parasypathomimetic agents, cholinesterase inhibitors, carbonicanhydrase inhibitors, and prostaglandins; combinations of anti-glaucomaagents; antioxidants; nutritional supplements; drugs for the treatmentof cystoid macular edema including, without limitation, non-steroidalanti-inflammatory agents; drugs for the treatment of ARMD, including,without limitation, angiogenesis inhibitors and nutritional supplements;drugs for the treatment of herpetic infections and CMV ocularinfections; drugs for the treatment of proliferative vitreoretinopathyincluding, without limitation, antimetabolites and fibrinolytics; woundmodulating agents, including, without limitation, growth factors;antimetabolites; neuroprotective drugs, including, without limitation,eliprodil; and angiostatic steroids for the treatment of diseases orconditions of the posterior segment of the eye, including, withoutlimitation, ARMD, CNV, retinopathies, retinitis, uveitis, macular edema,and glaucoma. Such angiostatic steroids are more fully disclosed in U.S.Pat. Nos. 5,679,666 and 5,770,592, which are incorporated herein intheir entirety by reference. Preferred ones of such angiostatic steroidsinclude 4,9(11)-Pregnadien-17α,21-diol-3,20-dione and4,9(11)-Pregnadien-17α,21-diol-3,20-dione-21-acetate. A preferrednon-steroidal anti-inflammatory for the treatment of cystoid macularedema is nepafenac.

A further embodiment of the subject invention provides for treatment ofproliferative and edematous diseases via concurrent administration of aCAI compound and an anti-angiogenic compound. Diverse anti-angiogeniccompounds can be used in a combinatorial treatment of the invention,include those with diverse mechanisms of action (e.g. VEGFneutralization, tyrosine receptor kinase inhibition, arachidonateinhibition, and Bcl-2 upregulation), such as glucocorticoids,non-steroidal anti-inflammatory drugs.

A further embodiment of the subject invention provides for the localadministration of CAI compound and related formulations thereof, incombination with other pharmacological therapies specifically designedto provide efficacy with limited side effects, e.g. neuroprotectantagents, inhibitors of drug efflux, metabolite inhibition, synergisticagents, and agents designed to decrease specific side effects.

A further embodiment of the subject invention provides for the treatmentof ocular diseases such as glaucoma and inflammatory eye disease byocular administration of CAI compound or formulations thereof, asdescribed herein in combination with other pharmacological agentsincluding, but are not limited to, verteporfin photodynamic therapy (QLTPharmaceuticals), anecortave acetate (Alcon Research Ltd.), Macugen®(Eyetech Pharmaceuticals), Lucentis® (ranibizumab, Genentech),Squalamine lactate (Genaera Corporation), LY333531 (Eli Lilly), andFluocinolone (Bausch & Lomb), timolol, bromodidine, cyclosporine,cis-platin, carboplatin, methotrexate, steroids, BDNF, CTF, and TNF-αblockers such as thalidomide and its derivatives and prodrugs.

Dexamethasone has been detected at 13 ng/ml in the vitreous cavityfollowing a single 5 mg peribulbar injection in humans (Weijtens et al.Ophthalmology 107(10): 1932-8 2000; Weijtens et al. Am J Ophthalmol128(2): 192-7 1999). This results in a vitreous concentration ofapproximately 0.10-0.13 μM assuming a vitreous volume of 4-5 ml.Delivery to the subretinal space is 10-fold higher followingsubconjunctival injection (Weijtens et al. Am J Ophthalmol 123(3):358-63 (1997)). The serum half-life of dexamethasone (18-36 hours) issignificantly shorter than that of CAI (111 hours). Physiological tissueconcentrations of CAI are in the 1-10 μM range so that it appearsreasonable to postulate based on the scleral permeability and prolongedserum half-life of CAI that it can be effectively deliveredtranssclerally to the subretinal space, and the vitreous cavity inphysiological concentrations (Weijtens et al. Ophthalmology 107(10):1932-8 2000; Weijtens et al. Am J Ophthalmol 128(2): 192-7 1999). Therelease rates of CAI compound or formulations thereof, fromsustained-release devices are also favorable. One potential drawback ofthe intravitreal implants that were constructed with CAI was that somedemonstrated rapid expansion because of the hygroscopic nature of CAI(Weijtens et al. Am J Ophthalmol 123(3): 358-63 1997). This rapidexpansion may be alleviated by developing a non-aqueous reservoir forthe CAI compound or formulations thereof, pellet, or by formulations ofa prodrug as stated in this patent.

The present invention also contemplates the use of a glucocorticoidand/or neuroprotective agent in combination with the CAI compound orformulations thereof. A glucocorticoid alone and/or neuroprotectiveagent in combination with CAI compound or formulations thereof, isuseful for treating persons suffering from pathologic ocularangiogenesis, in particular, exudative ARMD and/or PDR, as well assubretinal or retinal edema associated with either condition. Inaddition to being effective in inhibiting the neovascularizationassociated with wet ARMD and PDR, CAI compound or formulations thereof,could be useful in controlling any TOP rise associated with the use of aglucocorticoid, or to protect the retina from ischemic damage associatedwith microangiopathy or retinal vascular occlusions.

The present invention also contemplates the use of a glucocorticoidand/or neuroprotective agent in combination with the CAI compound (e.g.5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide)or formulations thereof for use in the treatment of uveitis, inparticular posterior infectious uveitis. For example, in one embodiment,the CAI compound may be used in combination with a glucocorticoid(exemplary glucocorticoids include, dexamethasone, fluoromethalone,administration of medrysone, budesonide, betamethasone, fluocinalone,triamcinolone, triamcinolone acetonide, prednisone, prednisolone,hydrocortisone, rimexolone, and pharmaceutically acceptable saltsthereof). In one embodiment, the CAI compound is used in combinationwith dexamethasone. In another embodiment, the CAI compound may be usedin combination with the glucorticoid and an anti-inflammatory agent.

D. Delivery Via Devices

A further embodiment provides the use of CAI compound or formulationsthereof, in conjunction with a drug delivery system in the form of animplant or a device for treatment of conditions as set forth herein.

The present invention also provides CAI compound or formulationsthereof, for use as coatings in conjunction with physical materialimplants such as stents and band ligatures used to treat vasculardiseases. In an embodiment, novel CAI compound or formulations thereof,coated stents of the subject invention are used in the treatment ofvascular disorders such as restenosis or vascular occlusion followingvascular insult (e.g., angioplasty, allo- or xenotransplantvasculopathies; variceal bleeding, and transplantation of an organ).

Delivery of drugs in the form of topical eye drops is also of littleutility when the drug is a protein or peptide that lacks the ability tocross the cornea and be made available to the vitreous, retina, or othersubretinal structures such as the retinal pigment epithelium (“RPE”) orchoroidal vasculature. An extraocular insert is a contact lens deliverysystem that releases medication over an extended period. The lensgenerally only lasts for a matter of hours or days before dissolving orreleasing all of the therapeutic compound. Continuous delivery ofmedication is inconvenient, requiring frequent re-application. Again,these contact lenses only provide drug to the cornea and anteriorchamber.

In rare cases, direct delivery of drugs has also been accomplished usingextemraliz tubes. This requires insertion of one end of a tube into thecorner of the patient's eye. The other end of the tube is taped to thepatient's forehead and terminates in a septum, through which medicationis delivered. This method is undesirable, being both uncomfortable andinconvenient. Since medication must be injected through the septum, thedevice is incapable of continuous delivery of medication. Furthermore,such tubes may become infected and in some cases ultimately threaten thepatient's vision. Direct delivery of drugs can also be accomplished bythe intraocular injection of the drug, or of microspheres that containor comprise the drug. However, microspheres tend to migrate within theeye, either into the visual axis or into adjacent tissue sites.

An intraocular insert is currently available for delivery of ganciclovirto the eye. Known as Vitrasert, the device consists of a nonerodible,polymer-based, sustained-release package containing ganciclovir, and anon-proteinaceous nucleoside analog. The device is surgically implantedin the vitreous humor of the eye to treat cytomegalovirus retinitis.See, e.g., Anand, R., et al., Arch. Ophthalmol., 111: 223-227 (1993).Another intraocular insert is disclosed by U.S. Pat. No. 5,466,233. Thistack-shaped device is surgically implanted so that the head of the tackis external to the eye, abutting the scleral surface. The post of thetack crosses the sclera and extends into the vitreous humor, where itprovides for vitreal drug release.

CAI compound or formulations thereof, of the invention may beadministered surgically as an ocular implant. As one example, areservoir container having a diffusible wall of polyvinyl alcohol orpolyvinyl acetate or partially hydrolyzed (e.g., about 1% to about 99%hydrolyzed) polyvinyl acetate and containing milligram (e.g., in therange of from about 1 mg to about 100 mg) quantities of CAI may beimplanted in or on the sclera. As another example, CAI in milligramquantities may be incorporated into a polymeric matrix having dimensionsof about 2 mm by 4 mm, and made of a biocompatible and optionallybiodegradable polymer such as polycaprolactone, poly(glycolic) acid,poly(lactic) acid, or a polyanhydride, or a lipid such as sebacic acid,and may be implanted on the sclera or in the eye. This is usuallyaccomplished with the patient receiving either a topical or localanesthetic and using a small (3-4 mm incision) made behind the cornea.The matrix, containing CAI compound or formulations thereof, is theninserted through the incision and sutured to the sclera using 9-0 nylon.

For example, U.S. Pat. No. 5,773,019, the contents of which is herebyincorporated by reference, discloses implantable controlled releasedevices for delivering drugs to the eye wherein the implantable devicehas an inner core containing an effective amount of a low solubilitydrug covered by a non-bioerodible polymer coating layer that ispermeable to the low solubility drug. U.S. Pat. No. 5,378,475, thecontents of which is hereby incorporated by reference, disclosessustained-release drug delivery devices that have an inner core orreservoir comprising a drug, a first coating layer which is essentiallyimpermeable to the passage of the drug, and a second coating layer whichis permeable to the drug. The first coating layer covers at least aportion of the inner core but at least a small portion of the inner coreis not coated with the first coating layer. The second coating layeressentially completely covers the first coating layer and the uncoatedportion of the inner core. U.S. Pat. No. 4,853,224, the contents ofwhich is hereby incorporated by reference, discloses biodegradableocular implants comprising microencapsulated drugs for implantation intothe anterior and/or posterior chambers of the eye.

The polymeric encapsulating agent or lipid encapsulating agent is theprimary element of the capsule. U.S. Pat. No. 5,164,188, the content ofwhich is hereby incorporated by reference, discloses the use ofbiodegradable implants in the suprachoroid of an eye. The implants aregenerally encapsulated. The capsule, for the most part, is a polymericencapsulating agent. Material capable of being placed in a given area ofthe suprachoroid without migration, “such as oxycel, gelatin, silicone,etc.” can also be used. U.S. Pat. No. 6,120,789, the contents of whichis hereby incorporated by reference, discloses the use of anon-polymeric composition for in situ formation of a solid matrix in ananimal, and use of the composition as a medical device or as asustained-release delivery system for a biologically-active agent, amongother uses. Such implants can provide CAI compound or formulationsthereof, of the invention to a patient to treat a non-life threateningdisease.

Another implantable device that can be used to deliver formulations ofthe present invention is the biodegradable implants described in U.S.Pat. No. 5,869,079, the contents of which is hereby incorporated byreference. Additional intracorporeal devices to which a CAI compound orformulations thereof, can be added for example in the form of a coatinginclude, but are not limited to, catheters, stents, angioplastyballoons, pacemakers, etc.

Within yet other aspects of the present invention, methods are providedfor expanding the lumen of a body passageway, comprising inserting astent into the passageway, the stent having a generally tubularstructure, the surface of the stent structure being coated with acomposition comprising a CAI compound, optionally in the presence ofpaclitaxel, such that the passageway is expanded. Within variousembodiments of the invention, methods are provided for eliminatingbiliary obstructions, comprising inserting a biliary stent into abiliary passageway; for eliminating urethral obstructions, comprisinginserting a urethral stent into a urethra; for eliminating esophagealobstructions, comprising inserting an esophageal stent into anesophagus; and for eliminating tracheal/bronchial obstructions,comprising inserting a tracheal/bronchial stent into the trachea orbronchi. Within each of these embodiments the stent has a generallytubular structure, the surface of the structure being coated with acomposition comprising a CAI compound or formulation thereof and,optionally, together with other pharmaceutically active agents for acombination therapy.

VI. ANIMAL EFFICACY AND SAFETY MODELS

The CAI parent compound, and the principles on which the CAI compound orformulations thereof, of the inventions described herein, have beendemonstrated to have antiproliferative and antimetastatic activity thatwas linked to decrease of intracellular calcium by inhibition ofnon-voltage-gated calcium channels. Furthermore, the anti-angiogenicactivity of CAI, the active ingredient contained in the CAI compound orformulations thereof, or applied via drug delivery systems and routes ofadministration that are the subject of this invention, conferringefficacy on neovascular disease has been well established.

The CAI compound or formulations thereof, used directly or inconjunction with a drug delivery method or device as described hereinhave highly advantageous therapeutic efficacy and safety for clinicaluse as local disease therapy when compared to systemic therapies of CAIdescribed to date. The advantages of the CAI compound or formulationsthereof, and administration methods of this invention are provided bythe controlled, efficacious, and safe free CAI drug concentrations tothe target tissue. The efficacy of CAI compound or formulations thereof,and drug delivery systems described herein to treat diseases describedherein can be confirmed using standard in vivo animal test model ofdisease. These models include the mouse retinal neovascularizationretinopathy of prematurity model (Smith L E et al. Invest Ophthalmol VisSci. 35(1):101-11, 1994, the method of which is hereby incorporated byreference), as well as the demonstration of an antiproliferative effecton choroidal endothelial cells and RPE in culture (Hoffman et al.Ophthalmologe 2004, the method of which is hereby incorporated byreference). Further, the ability of CAI compound or formulationsthereof, to provide controlled, safe levels suitable to treat humanproliferative or neovascular ocular diseases can be confirmed usingstandard in vivo animal test ocular pharmacokinetic and pharmacodynamicstudies in animals.

The CAI parent compound active ingredient from which the CAI compound orformulations thereof, inventions described herein derive, hasdemonstrated primary pharmacological action to decrease intracellularcalcium concentration by inhibition of non-voltage-gated calciumchannels (Kohn, E. C. et al. CAI. Cancer Res. 54:935-942, 1994) thusaffecting diverse signal transduction processes. Furthermore, thepharmacological mechanisms of action of CAI include (but are not limitedto) the inhibition of disease mediating molecules including growthfactors (e.g. VEGF), cytokines (e.g. IL-6), matrix metalloproteinases(MMps) and arachadonic acid (Felder, C. C. et al. J Pharmacol Exp Ther.257:967-971, 1991; Fox D A et al. Ann N Y Acad Sci.; 893:282-5, 1999;Cole K. et al. Cancer Metastasis Rev. March; 13(1):31-44, 1994). Thesepharmacological mechanisms described for CAI provide the fundamentalbasis of anti-proliferative activity on diverse cancer cell types(melanoma, breast, squamous, prostate, ovarian, glioblastoma, colon, andsmall lung cell cancer) relevant to current clinical application of CAIas a systemically administered drug in life-threatening cancerindications. Furthermore, these pharmacological mechanisms described forCAI provide the fundamental basis of anti-proliferative activity againstother non-cancer cell types directly relevant to the invention,including but not limited to, human vascular and retinal endothelialcells, retinal pigment and choroidal endothelial cells (Hoffman et al.Ophthalmologe 2004), inflammatory cells including lymphocytes andeosinophils. Furthermore, since pathological increases in intracellularcalcium (e.g. as mediated by pathological activation of NMDA receptorchannels by excess glutamate in ischemic conditions) in neurons is knownto result in neuronal death, it is anticipated by this invention thatadministration of CAI compound or formulations thereof, as describedherein can be neuroprotective. Therefore, based on the pharmacologicalmechanisms CAI compound or formulations thereof, described herein haveanti-angiogenic, anti-inflammatory and neuroprotective effectsassociated with the treatment of diseases of relevant diseases asdescribed herein.

The CAI compound or formulations thereof, used directly or inconjunction with a drug delivery method or device as described hereinhave highly advantageous therapeutic efficacy and safety for clinicaluse as local disease therapy when compared to systemic therapies of CAIdescribed to date. The advantages of the CAI compound or formulationsthereof, and administration methods of this invention are provided bythe controlled, efficacious, and safe free CAI drug concentrationsadministered to or reaching the target tissue. Plasma concentrations ofCAI that have been associated with efficacy in cancer described inpatients range from approximately 1.0 to 10 micromolar while plasmaconcentrations of above 100 micromolar can be associated withunacceptable toxicity. The invention anticipates pharmacologicallyactive concentrations of CAI as provided by the CAI compound orformulations thereof, of this invention to exhibit therapeutic effectsin the local target tissue to range between 0.5 and 100 micro molar. Asa specific example this invention anticipates periocular administrationof a CAI compound or formulations thereof, which formulations contain 5mg of active CAI ingredient, to provide CAI to the target subretinal andvitreal compartments in concentrations ranging from 0.1 and 10micromolar concentration over a one-day time period. The ability of CAIcompound or formulations thereof, to provide controlled, safe levelssuitable to diseases described herein can be confirmed using standard invivo pharmacokinetic studies in healthy animals or animal diseasemodels. The CAI compound or formulations thereof, is administered asdescribed herein and the samples of the target tissue are analyzed forconcentration of CAI active principle using known e.g., (Tutsch, K. D.et al. Proc Am Assoc Cancer Res. 37:A1133, 1996, the contents of whichare hereby incorporated by reference in their entirety) or tissueadapted bioanalytical methods. For example, a CAI formulation containing5 mg of active CAI ingredient is given to rabbits by periocularadministration, and the animals are sacrificed at different time pointsover a 24 hour period. Tissue and liquid humor samples are taken fromthe eyes and the CAI active ingredient is extracted and subjected toHPLC-MS analysis with an appropriate internal standard to the targetdisease compartments including retinal, choroidal and vitrealcompartments.

The efficacy of CAI compound or formulations thereof, alone or inconjunction with drug delivery systems described herein to treatdiseases described herein can be confirmed using in vivo animal testmodels of disease. These models include the mouse retinalneovascularization retinopathy of prematurity model (Smith L E et al.Invest Ophthalmol Vis Sci. 35(1):101-11, 1994, the methods of which arehereby incorporated by reference). In this model, mouse pups are placedinto 75% 02, along with their nursing dams, at post-natal day 7. Theyare maintained at this oxygen level for five days, at which point theyare returned to normoxia. For the next five days, the pups areadministered with CAI compound or formulations thereof, using themethods described herein. At day 17, the animals are euthanized,enucleated, and the eyes fixed in 4% buffered paraformaldehydeovernight, then transferred to saline, embedded in paraffin, andsectioned. Serial sections are collected (6 micron sections, every30^(th) section) and stained with hematoxylin and eosin. Individualsmasked to the identity of treatment then count the number of pre-retinalnuclei and the effects of CAI compound or formulations thereof, onlowering the number of pre-retinal nuclei are evaluated relative tocontrol. At least eight sections are counted for each eye. Another modelis the adult mouse model of ischemia-induced choroidalneovascularization that mimics age-related macular degeneration. In thismodel, adult mice are subjected to laser photocoagulation in a mannersimilar to Ryan's (Ryan S J. et al. Trans Am Ophthalmol Soc. 77:707-745,1979, the methods of which are hereby incorporated by reference). Threeburn spots are produced in the choroid using an argon green wavelengthlaser at a power of 910-1030 mW for 0.05 sec to induce choroidal ruptureand subsequent neovascularization. The burns are in three quadrants ofthe choroid, one disc area in diameter and one disc area from the opticnerve. This fixed distance allows the burns to be both reproducible andisolated from each other. The laser produces a bubble in the majority ofcases, which is indicative of a Bruch's membrane rupture. In less than5% of the animals, bleeding may be noted upon treatment. The mice areeuthanized at two weeks after receiving laser photocoagulation and theeyes removed for evaluation of choroidal neovascularization.

Other disease models include xenograft models for host vs. graft tissuerejection relevant to applications including restenosis. Another objectof the present invention is to provide for local treatment of otherproliferative and angiogenic diseases including topical delivery of CAIcompound or formulations thereof, for severe dermatological diseasesincluding severe psoriasis, excema and rosacea and local intrarticularadministration for severe arthritis by inhibiting vascular andinflammatory cell proliferation.

VII. THERAPEUTIC APPLICATIONS

In one embodiment, the CAI compound and formulations thereof of thepresent invention can be used for administration to a human patientsuffering from a non-life threatening disease, as a method of treatmentof such disease. The non-life threatening disease can be, for example aproliferative disease, an inflammatory disease, an edematous disease, aneurodegenerative and/or a neurotoxic disease, or a signaltransduction-mediated disease, or a matrix matalloproteinase-mediateddisease.

Examples of diseases or disorders that can be mediated directly, orindirectly, by the administration of a CAI compound or formulationsthereof, of the invention include, but are not limited to: age-relatedmacular degeneration, diabetic retinopathy, retinal vascular occlusion,choroidal and retinal angiomatous proliferation, chronic glaucoma,retinal detachment, sickle cell retinopathy, rubeosis iridis, uveitis,neoplasms, Fuchs heterochromic iridocyclitis, neovascular glaucoma,corneal neovascularization, neovascularization resulting from combinedvitrectomy and lensectomy, retinal ischemia, choroidal vascularinsufficiency, choroidal thrombosis, carotid artery ischemia, retinalartery/vein occlusion, e.g., central retinal artery occlusion and branchretinal vein occlusion, contusive ocular injury, and retinopathy ofprematurity, and other vascular anomalies, e.g., retinitis pigmentosa,endophthalmitis, infectious diseases, inflammatory but non-infectiousdiseases, ocular ischemia syndrome, peripheral retinal degenerations,retinal degenerations and tumors, choroidal disorders and tumors,vitreous disorders, retinal detachment, non-penetrating and penetratingtrauma, post-cataract complications, and inflammatory opticneuropathies.

In one embodiment, the disease that can be mediated directly, orindirectly, by the administration of a CAI compound or formulationsthereof may be uveitis or any other inflammation of the uvea. Inparticular, the CAI compound or formulations may be used to treatinfectious uveitis. In one embodiment, the CAI compound may be used totreat infectious posterior uveitis (e.g. inflammation of the retina orchoroid). In certain embodiments, the infectious posterior uveitis maybe due to parasites or bacteria. For example, infectious uveitis mayencompass inflammation of the uvea associated with one or more oftoxoplasmosis, bartonellosis, histoplasmosis, Lyme disease, syphilis,and tuberculosis.

Hereditary degenerative retinal and vitreoretinal diseases treatablewith a CAI compound or formulations thereof, of the invention, eitheralone or in combination therapies, include: Primary pigmentedretinopathies, all gene types (ocular involvement only); Autosomaldominant retinitis pigmentosa e.g. rod-cone and cone-rod degenerations;Autosomal recessive retinitis pigmentosa e.g. rod-cone and cone-rodedegenerations, Leber's amaurosis congenita; X-linked recessive pigmentedretinopathies e.g. choroideremia; Secondary pigmented retinopathies(retinopathies associated with systemic diseases); Autosomal dominantpigmented retinopathies, e.g. Paget's disease, Charcot-Marie-Toothdisease, Steinert's disease, Pierre-Marie syndrome; Autosomal dominantpigmented retinopathies e.g. diabetes mellitus, mannosidoses,mucopolysccharidoses, Batten's disease, Refsum's disease, Ushersyndrome; X-linked recessive pigmented retinopathies e.g. Huntersyndrome; conjunctivitis (e.g. allergic conjunctivitis, chronicconjunctivitis, contact lens-associated conjunctivitis, conjunctivalulceration, drug-related conjunctivitis); uveitis, uveoretinitis,chronic diseases (e.g. age-related macular panuveitis, retinitis,degeneration diabetes mellitus, infectious choroiditis, vitreitis,diseases (e.g., tuberculosis syphilis,cytomegalo-Scleritis/Episcleritis, virus retinitis), injury as a resultof physical agents (e.g. UV Iridocyclitis, radiation), chemical agents(e.g. acids, Endophthalmitis, caustic solvents), and immunologicaletiologies (e.g. sarcoidosis, inflammatory bowel disease, Cornealulceration disease, and other collagen vascular diseases). VonHippel-Lindau syndrome is a specific neovascular disease that has bothocular and non-ocular manifestations that should be treatable by CAIcompounds and formulations thereof.

The compounds, formulations, and methods of the present invention mayalso be used to treat ocular symptoms resulting from diseases orconditions that have both ocular and non-ocular symptoms. Some examplesinclude AIDS-related disorders such as cytomegalovirus retinitis anddisorders of the vitreous; pregnancy-related disorders such ashypertensive changes in the retina; and ocular effects of variousinfectious diseases such as tuberculosis, syphilis, lyme disease,parasitic disease, toxocara canis, ophthalmonyiasis, cyst cercosis, andfungal infections. Non-ocular diseases can also include rheumatoidarthritis, psoriasis, contact dermatitis, keratitis, conjunctivitis,scleritis, squamous cell carcinoma and condyloma.

Angiogenesis and neovascularization in the adult animal is usually apathological process, and is in direct contradistinction tonon-pathological neovascularization, which usually occurs in normalembryogenesis (e.g., development of the embryonic vascular system). Inaccordance with the subject invention, neovascularization refersspecifically to pathological neovascularization. Aberrant orpathological vascularization is a key component in numerous diseasestates. For example, vascularization is a critical element of most solidtumors, such as cancers of the brain, genitourinary tract, lymphaticsystem, stomach, larynx and lung. These include histiocytic lymphoma,lung adenocarcinoma and small cell lung cancers. Aberrant vasculargrowth in the retina can lead to visual degeneration, which canculminate in blindness. Accordingly, the subject invention provides CAIcompound and formulations thereof for the treatment ofneovascularization.

Compounds of the invention can also be used to inhibit the proliferationof vascular endothelial cells and so are indicated for use in treatinggraft vessel diseases such as restenosis or vascular occlusion followingvascular insult such as angioplasty, allo- or xenotransplantvasculopathies, graft vessel atherosclerosis, and in the transplantationof an organ (e.g., heart, liver, lung, kidney or pancreatic transplants(Weckbecker et al., Transplantation Proceedings 1997, 29, 2599-2600).

The present invention also provides for the treatment of severedermatological diseases including severe psoriasis, contact dermatitis,excema and rosacea; severe arthritis; and other vascular andinflammatory cell proliferative diseases (such as rheumatoid arthritis;spondyloarthropathies; gouty arthritis; osteoarthritis; systemic lupuserythematosus; juvenile arthritis; asthma; bronchitis; menstrual cramps;tendonitis; bursitis; skin related conditions such as psoriasis, eczema,burns, and dermatitis; gastrointestinal conditions such as inflammatorybowel disease, Crohn's disease, gastritis, irritable bowel syndrome andulcerative colitis; colorectal cancer; migraine headaches; periarteritisnodosa; thyroiditis; aplastic anemia; Hodgkin's disease; sclerodoma;rheumatic fever; type I diabetes; myasthenia gravis; sarcoidosis;nephrotic syndrome; Behcet's syndrome; polymyositis; gingivitis;hypersensitivity; swelling occurring after injury; myocardial ischemia;and the like).

Additionally, disease states which rely on aberrant signaltransduction/proliferation may also be treated by the CAI compound orformulations thereof, or methods of the invention. Diseases ofpotentially aberrant signal transduction/proliferation may include thecollagen vasculitides (i.e., systemic lupus erthythematosis andrheumatoid arthritis), neurologic diseases (i.e., dementia and nerveconduction diseases), diseases of transport (i.e., cystic fibrosis),toxic effects of agents (i.e., cisplatin-related neuropathy), andcellular dysfunction (i.e., myelodysfunction syndromes), hemangiomata,and collagen vasculidities.

One embodiment of the subject invention provides for the local ocularadministration of CAI compound or formulations thereof, by means ofperiocular, retrobulbar, intravitreal, subretinal, posteriorjuxtascleral, topical and subconjunctival administration throughinjection or needle-free system, or topical administration though localinstillation, drops, ointment, or in conjunction with drug deliverysystems exemplified by contact lenses, devices and implants. A relatedembodiment provides the treatment of a broad range of ocular diseasesincluding diabetic retinopathy (DR), neovascular proliferativeage-related macular degeneration (ARMD), diabetic macular edema (DME),cystoid macular edema (CME) and ocular tumors such as retinoblastoma(RB), Retinopathy of Prematurity (ROP), retinal vascular occlusions(RVO), uveitis, glaucoma, corneal neovascularization, irisneovascularization, neovascular glaucoma, ischemic neural damage, andpterygium by the local ocular administration of CAI compound and relatedformulations thereof, directly or in conjunction with a drug deliverysystem as described herein.

Following are examples, which illustrate procedures for practicing theinvention. These examples should not be construed as limiting.

It is advantageous to define several abbreviations before describing theexamples. It should be appreciated that the following abbreviations areused throughout this application.

CAI=5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamideTween® 80=polyoxyethylenesorbitan monooleatePluronic®=polyoxyethylene-polyoxypropylene block copolymerPluronic® F-68,F-127=α-hydro-omega-hydroxypoly(oxyethylene)-poly(oxypropylene)-poloxyethyleneblock copolymers

Example 1 Procedures for Preparing and Identifying Stable, AutoclavableAqueous Formulations of CAI Compounds General Procedure

Amorphous solid free base form of CAI active pharmaceutical ingredienthaving Formula I(5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide),which can be synthesized by one skilled in the art of organic chemistryaccording to procedures described in U.S. Pat. Nos. 4,590,201 and5,602,156, is first dissolved in a minimal volume of absolute ethanol at40° C. Additives are added to this solution as selected from an ionicsurfactant (preferably a lipophilic organic acid having at least 8carbon atoms such as oleic acid), an organic oil, such as alkyl ester offatty acid and/or monoglyceride, diglyceride, or triglyceride, orglyceride (e.g. soybean oil, or mixtures thereof), and other organicsolubilizing additives (e.g. Tweens, Pluronics, cyclodextrins,polysorbates, sorbitan esters, sucroesters, and other non-ionicsurfactants, polyethylene glycols, particularly those that are liquid atambient temperature) in proportions that result in a clear solutioncontaining CAI in concentrations from 1 to 200 mg/mL. Preferredproportions of these primary formulation ingredients are selected basedon suitability for achieving concentrations of CAI from 1 to 100 mg/mL.After adding water to the resulting organic solution of CAI a stableorganic-aqueous mixture comprising CAI results as a homogeneoussolution, stable emulsion, or self-emulsifying dry powder. The ethanolis removed from the resulting organic mixture described above byevaporation (e.g. under reduced pressure using a rotary evaporator).Water is added to the resulting organic mixture, and the pH is adjustedto an acceptable therapeutically applicable range (preferably from aboutpH 3.0 to about pH 8.0 and more preferably from about pH 5.0 to about pH8.0). The exact pH adjustment is done by addition of a basic solution(preferably aqueous sodium hydroxide) or an acidic solution (preferablyaqueous hydrochloric acid) or both in quantities sufficient to providethe desired pH.

Preliminary preferred formulations are first selected by simpleinspection based on stability of the final solution against separationinto distinct organic and aqueous phases after standing for at least 3days within an ambient temperature range. Formulations that phaseseparate are unstable and are discarded. Stable sterile formulations arethen prepared by autoclaving for sufficient time at temperaturessuitable for effectively killing the microorganisms that may be presentin the formulation, for instance, 121° C. for about 30 minutes.

An alternate method for manufacturing stable sterile formulations can bereadily accomplished by filtration through sterile filtration membranes(e.g., 0.1 micron membranes, 0.2 micron membranes).

Formulation 1

Formulation 1 is prepared according to the General Procedure describedin Example 1 using the following amounts of reagents or correspondingrelative proportions: CAI free base (50 mg), oleic acid (150 mg),soybean oil (150 mg), Tween®-80 (100 mg), Pluronic® F-68 (100 mg). Acreamy emulsion of milky white appearance is formed after dilution withwater to a final CAI concentration of 36 mg/mL (3.6%). The emulsion ofFormulation 1 is stable based on stability to phase separation asdefined in Example 1. The active CAI ingredient is stable to autoclavesterilization conditions of 121° C. at a pressure of 15 psi as definedin Example 1. Thus, after autoclaving Formulation 1 under theseconditions for 30 min no decomposition of CAI or formation of newimpurities could be detected by reverse phase high pressure liquidchromatography (RP-HPLC) analysis using UV detection (Conditions: RP-18column: 5μ particle size, length=13 cm, diameter=8 mm; solvent system:solvent A=0.001% trifluoroacetic acid, solvent B=acetonitrile,gradient=80% A/20% B to 10% A/90% B over 12 min at a flow rate of 2.5mL/min; CAI retention time=6.3 min).

Formulation 2

Formulation 2 is prepared according to the General Procedure describedin Example 1 using the following amounts of reagents or correspondingrelative proportions: CAI free base (50 mg), oleic acid (200 mg),soybean oil (200 mg) and Tween®-80 (100 mg). An emulsion of milky whiteappearance is formed after dilution with water to a final CAIconcentration of 60 mg/mL (6.0%). The emulsion of Formulation 2 isstable based on stability to phase separation at ambient temperature.The active CAI ingredient is stable to autoclave sterilizationconditions of 121° C. at a pressure of 15 psi as defined in Example 1.Thus, after autoclaving Formulation 2 under these conditions for 30 minno decomposition of CAI or formation of new impurities could be detectedby RP-HPLC analysis using UV detection (Conditions: RP-18 column: 5μparticle size, length=13 cm, diameter=8 mm; solvent system: solventA=0.001% trifluoroacetic acid, solvent B=acetonitrile, gradient=80%A/20% B to 10% A/90% B over 12 min at a flow rate of 2.5 mL/min; CAIretention time=6.3 min).

Formulation 3

Formulation 3 is prepared according to the General Procedure describedin Example 1 using the following amounts of reagents or correspondingrelative proportions: CAI free base (50 mg), oleic acid (300 mg),Pluronic® F-127 (100 mg) and Tween®-80 (100 mg). A creamy emulsion ofmilky white appearance is formed after dilution with water to a finalCAI concentration of 36 mg/mL (3.6%). The emulsion of Formulation 3 isstable based on stability to phase separation at ambient temperature.The active CAI ingredient is stable to autoclave sterilizationconditions of 121° C. at a pressure of 15 psi as defined in Example 1.Thus, after autoclaving Formulation 3 under these conditions for 30 minno decomposition of CAI or formation of new impurities could be detectedby RP-HPLC analysis using UV detection (Conditions: RP-18 column: 5μparticle size, length=13 cm, diameter=8 mm; solvent system: solventA=0.001% trifluoroacetic acid, solvent B=acetonitrile, gradient=80%A/20% B to 10% A/90% B over 12 min at a flow rate of 2.5 mL/min; CAIretention time=6.3 min).

Formulation 4

Formulation 4 is prepared according to the General Procedure describedin Example 1 using the following amounts of reagents or correspondingrelative proportions: CAI free base (50 mg), oleic acid (150 mg),soybean oil (150 mg), Pluronic® F-68 (100 mg) and Tween®-80 (100 mg). Acreamy emulsion of milky white appearance is formed after dilution withwater to a final CAI concentration of 36 mg/mL (3.6%). The emulsion ofFormulation 4 is stable based on stability to phase separation atambient temperature. The active CAI ingredient is stable to autoclavesterilization conditions of 121° C. at a pressure of 15 psi as definedin Example 1. Thus, after autoclaving Formulation 4 under theseconditions for up to 30 min no decomposition of CAI or formation of newimpurities could be detected by RP-HPLC analysis using UV detection(Conditions: RP-18 column: 5μ particle size, length=13 cm, diameter=8mm; solvent system: solvent A=0.001% trifluoroacetic acid, solventB=acetonitrile, gradient=80% A/20% B to 10% A/90% B over 12 min at aflow rate of 2.5 mL/min; CAI retention time=6.3 min).

Formulation 5

Formulation 5 is prepared according to the General Procedure describedin Example 1 using the following amounts of reagents or correspondingrelative proportions: CAI free base (50 mg), oleic acid (150 mg),soybean oil (150 mg), Pluronic® F-127 (100 mg) and Tween®-80 (100 mg). Acreamy emulsion of milky white appearance is formed after dilution withwater to a final CAI concentration of 36 mg/mL (3.6%). The emulsion ofFormulation 4 is stable based on stability to phase separation atambient temperature. The active CAI ingredient is stable to autoclavesterilization conditions of 121° C. at a pressure of 15 psi as definedin Example 1. Thus, after autoclaving Formulation 5 under theseconditions for 30 min no decomposition of CAI or formation of newimpurities could be detected by RP-HPLC analysis using UV detection(Conditions: RP-18 column: 5μ particle size, length=13 cm, diameter=8mm; solvent system: solvent A=0.001% trifluoroacetic acid, solventB=acetonitrile, gradient=80% A/20% B to 10% A/90% B over 12 min at aflow rate of 2.5 mL/min; CAI retention time=6.3 min).

Formulation 6

Formulation 6 is prepared according to the General Procedure describedin Example 1 using the following amounts of reagents or correspondingrelative proportions: CAI free base (25 mg), oleic acid (150 mg),soybean oil (150 mg), Pluronic® F-68 (100 mg) and Tween®-80 (100 mg). Acreamy emulsion of milky white appearance is formed after dilution withwater to a final CAI concentration of 8 mg/mL (0.8%). The emulsion ofFormulation 6 is stable based on stability to phase separation atambient temperature. The active CAI ingredient is stable to autoclavesterilization conditions of 121° C. at a pressure of 15 psi as definedin Example 1. Thus, after autoclaving Formulation 6 under theseconditions for 30 min no decomposition of CAI or formation of newimpurities could be detected by RP-HPLC analysis using UV detection(Conditions: RP-18 column: 5μ particle size, length=13 cm, diameter=8mm; solvent system: solvent A=0.001% trifluoroacetic acid, solventB=acetonitrile, gradient=80% A/20% B to 10% A/90% B over 12 min at aflow rate of 2.5 mL/min; CAI retention time=6.3 min).

Example 2 Intravitreal Injection

Pharmacologically effective concentrations in diverse in vitro and invivo model systems of CAI at 0.5 to 10 micromolar concentration which isconsistent with a dose of 0.2 ng to 40 ng of drug in a compartment ofabout 4 to 5 mL for the eye for a long term 4 to 6 week maintenance of acontrolled dose use with indicate a dose range of 50 ng to 500 ng of CAIactive ingredient or greater.

The intravitreal injection would be administered via the inferotemporalpars plana, in a protocol similar to triamcinalone, Macugen®, orLucentis®. As a single dose of 5 mg of dexamethasone-administeredsubconjunctivally can reach micromolar concentrations in the subretinalspace and vitreous, and CAI possesses similar trans-scleral permeabilitykinetics, an initial dose of 5 to 50 mg for periocular administration isproposed. An effective quantity of the compound of interest is employedin treatment.

As understood by the skilled clinician, the dosage of compounds used inaccordance with the invention varies depending on the compound and thecondition being treated. For example, the age, weight, and clinicalcondition of the recipient patient, and the experience and judgment ofthe clinician or practitioner administering the therapy are among thefactors affecting the selected dosage. Other factors include the routeof administration, the patient, the patient's medical history, theseverity of the disease process, and the potency of the particularcompound. The dose should be sufficient to ameliorate symptoms or signsof the disease treated without producing unacceptable toxicity to thepatient. In general, an effective amount of the compound is that whichprovides either subjective relief of symptoms or an objectivelyidentifiable improvement as noted by the clinician or other qualifiedobserver.

Example 3 Single Ingredient Formulation

Ingredient Concentration w/v % CAI 0.1 to 2% Monobasic Sodium PhosphateDihydrate 0.051% Dibasic Sodium Phosphate Dodecahydrate 0.5% Tyloxapol0.05-0.4% Sodium Chloride 0.76% NaOH/HCl pH adjust to 5.0-8.4 Water forinjection q.s. 100%.

Example 4 Steroid Combination Formulation

Ingredient Concentration w/v % CAI 0.1 to 2% Triamcinolone Acetonide0.5-4.0% Monobasic Sodium Phosphate Dihydrate 0.051% Dibasic SodiumPhosphate Dodecahydrate 0.5% Tyloxapol 0.05-0.4% Sodium Chloride 0.76%NaOH/HCl pH adjust to 5.0-8.4 Water for injection q.s. 100%.

Example 5 Neuroprotective Combination Formulation

Ingredient Concentration w/v % CAI 0.1 to 2% Neuroprotecive agent0.5-4.0% Monobasic Sodium Phosphate Dihydrate 0.051% Dibasic SodiumPhosphate Dodecahydrate 0.5% Tyloxapol 0.05-0.4% Sodium Chloride 0.76%NaOH/HCl pH adjust to 5.0-8.4 Water for injection q.s. 100%.

Example 6 Chelating Agent Combination Formulation

Ingredient Concentration w/v % CAI 0.1 to 2% Chelating Agent 0.5-4.0%Monobasic Sodium Phosphate Dihydrate 0.051% Dibasic Sodium PhosphateDodecahydrate 0.5% Tyloxapol 0.05-0.4% Sodium Chloride 0.76% NaOH/HCl pHadjust to 5.0-8.4 Water for injection q.s. 100%.

Example 7 Topical Formulation

A typical example of topical formulation of CAI is as follows:Ingredient Concentration w/v % (Preferred Range) CAI Polyquad0.0005-0.01% (0.0001%); HPMC (hydroxypropylmethylcellullose) 0.02-1.0%(0.5%); Mannitol (b) 0.0-5.0% (3.82%); Sodium Chloride (d) 0.0-0.8%(0.17%); Disodium Edetate 0.0-0.2% (0.01%); Polysorbate-80 (c)0.005-0.4% (0.05%); NaOH and/or HCl q.s. pH 5.0-8.4 (6.8-7.8); PurifiedWater q.s. 100%. (a) Other suitable polymers include cellulosic polymerslike HPMC, HEC (hydroxyethylcellulose), sodium CMC (sodiumcarboxymethylcellulose), polyvinyl alcohol (PVA), Polyvinyl Pyrrolidone(PVP), polyacrylamide, and other water miscible/soluble polymers toimpart viscosity to the product and to stabilize suspension. (b) Bothionic as well nonionic agents are used to adjust Osmolality of theproduct either alone or in combination. This also stabilizes thesuspension. (c) Other surfactants that can be used are non-ionic(Tyloxapol, Tweens, Spans) anionic (lecithin, hydrogenated lecithins),or anionic (sodium lauryl sulfate, bile salts).

The following contemplated clinical cases of angiogenesis are presentedas examples of methods of treatment and not as limitations. Otherillustrations of treating systemic or localized angiogenesis are notpresented, since the principles involved are similar, and will beapparent to the skilled reader based upon review of an individual'sclinical manifestation and the preferred embodiments of the inventivecompositions.

Example 8 Diabetic Complications

Patient A is determined to be experiencing an acute relapse of diabetes,e.g., diabetic coma. Patient A has a history of diabetes over the past20 years, with increasing debilitation from progressive diabeticretinopathy, nephropathy, and neuropathy. Patient A has not yet hadpanretinal laser photocoagulopathy but has been treated with focal laserfor macular edema. Retinal examination reveals Patient A has notprogressed to threshold criteria for panretinal photocoagulation per theDiabetic Retinopathy Study (DRS) guidelines.

According to the subject invention, Patient A will require scheduling ofa follow-up visit and low-dose treatment with a CAI compound of theinvention in an attempt to delay or remove the need for panretinal laserphotocoagulopathy. Patient A is given a direct intravitreal injection ofa high dose of CAI, 50 microgram/injection every week for 4 weeks. Thedose remains constant with weekly injections and Patient A is scheduledfor panretinal laser photocoagulopathy with continued weekly retinalexamination follow-up to determine disease course and confirm necessityfor panretinal laser photocoagulopathy.

After regression of the neovascularization, Patient A is treated withfocal laser therapy for macular edema, if needed, and provided a withmonthly, or less frequent, intravitreal injections of CAI with theadministration frequency and dose adjusted according to patientsclinical manifestations and to maintain neovascular regression.Intravitreal injections may be supplemented or replaced by otherappropriate routes of administration and this could be dictated by thepatients clinical presentation, health, expected treatment complianceprofile, and appropriateness of local, systemic, or controlled releasetreatment. CAI treatment would be targeted to be tapered off toprophylactic systemic or controlled release therapy, as the conditionimproves sufficiently.

Patient B is also determined to be experiencing an acute relapse ofdiabetes, e.g., diabetic coma and has a history of diabetes over thepast 20 years, with increasing debilitation from progressive diabeticretinopathy, nephropathy, and neuropathy. Patient B has not yet hadpanretinal laser photocoagulopathy nor has she been treated with focallaser for macular edema. Patient B has not developed thresholdproliferative diabetic retinopathy. This patient is clinically similarto Patient A, but the evidence of no macula involvement obviates theneed for scheduling focal laser photocoagulation.

According to the subject invention, Patient B is given a directintravitreal injection of a low dose of CAI, 5 microgram/injection. Thedose remains constant with a second intravitreal injection 4 weekslater. Patient B is followed clinically every 3 months and treated withfocal laser therapy, if needed, or provided with subsequent intravitrealinjections of CAI with the administration frequency and dose adjustedaccording to Patient B's clinical manifestations. Intravitrealinjections may be supplemented or replaced by other appropriate routesof administration and this could be dictated by the patients clinicalpresentation, health, expected treatment compliance profile, andappropriateness of local, systemic, or controlled release treatment. CAItreatment is targeted as prophylactic systemic or controlled releasetherapy, if the condition improves sufficiently.

Example 9 Proliferative Diabetic Retinopathy

62-year old male with Proliferative Diabetic Retinopathy and thresholdhigh-risk characteristics presents with Vitreous Hemorrhage. The patientalso has atrial fibrillation and ischemic cardiomyopathy that requiresoral Coumadin. The patient is intolerant to topical PanretinalPhotocoagulation. The INR measured at just less than 2. Because of thisacute event intravitreal injection, a medium dose of 50 mcg CAI isadministered with some neovascular regression. The patients VitreousHemorrhage slowly resolves and the patient is able to taper his Coumadinso that Panretinal Photocoagulation can be placed with retrobulbaranesthesia. Alternately, the patient remains on Coumadin and then istreated with a patch periocular insert of CAI to diminish theneovascularization.

Example 10 Central Scotoma Treatment Complications

78-year old woman with a two-week history of central scotoma. Shedeveloped a large disciform scar in the contralateral eye after severalsessions of photodynamic therapy. On exam and fluorescein angiography ispredominately classic subfoveal membrane is noted. Intravitrealinjection of a high dose, 200 mcg, CAI is given to hasten theneovascular regression. After this high dose of intravitreal CAI aperiocular injection of CAI in a sustained-release solution is given andneovascular regression occurs.

Example 11 Age-Related Macular Degeneration Complication

67-year old white male who has had three years of poor vision secondaryto a disciform scar associated with Age-Related Macular Degeneration. Hehas high-risk drusen and parafoveal pigmentation. A long acting low dosesubconjunctival or periocular implant of CAI 1 mg is affixed to thewell-sighted eye to prophylax against neovascularization.

Example 12 Glaucoma Complication

82-year old woman with low-tension glaucoma. She has a history ofhypotony maculopathy in the left eye after trabeculectomy with visualacuity loss to 20/200 secondary to hypotony maculopathy associated withan intraocular pressure of 7 to 8. In the right eye, she isprogressively cupping with a Cup/Disc ratio 0.85 despite maximal medicaltreatment and an intraocular pressure of 11. A low dose 1 mg periocularinsert for injection of CAI is given in order to maximizeneuroprotection.

Example 13 Proliferative Diabetic Retinopathy

28-year old noncompliant diabetic male who initially presented withaggressive Proliferative Diabetic Retinopathy. Heavy and full PanretinalPhotocoagulation was performed, however there is recurrent residualVitreous Hemorrhaging from a small active frond of neovascularizationthat is within heavy Panretinal Photocoagulation scars. An intravitrealinjection of moderate dose, 100 mcg of CAI is given to cause neovascularinvolution and the patient is maintained with a moderate dose of 10 mgadministered by periocular injection of CAI in a sustained-releasesolution.

Example 14 Proliferative Diabetic Retinopathy

37-year old white male who is with Proliferative Diabetic Retinopathyand has undergone Trans Pars Plana Vitrectomy x's 2 for VitreousHemorrhage. He has a heavy pattern of Panretinal Photocoagulation scarsand it is thought that he has neovascularization arising from thesclerotomy sites, and/or the pars plana, which is causing persistenthemorrhage. A high dose of 200-mcg dose of CAI is administeredintravitreally and then two weeks later an outpatient air-fluid exchangeis performed, and a moderate dose, periocular injection of 10 mg of CAIis given to prevent further Vitreous Hemorrhage.

Example 15 Vision Loss

16-year old female who presents with a several month history of visionloss and is found to have a large angiomatous lesion that is causing asubtotal retinal detachment. Secondary to the lesion size it is notamenable for cryotherapy and standard laser photocoagulation. Initiallya high dose of 200 mcg CAI is given intravitreally and two weeks later aperiocular insert is placed of 10 mg, moderate dose of CAI onto thesclera overlying the lesion.

Example 16 Central Retinal Vein Occlusion

66-year old male who presents with a Central Retinal Vein Occlusion andflorid Iris Neovascularization with Neovascular Glaucoma. As an adjunctto Panretinal Photocoagulation secondary to the florid neovascularactivity, a high dose of 200 mcg CAI is given as a synergistic measureto cause regression of the anterior segment neovascularization andnormalization of the pressure.

Example 17 Iris and Preretinal Neovascularization

39-year old white male strikes his head on a laboratory cabinet. Thiscauses a chronic peripheral retinal detachment with irisneovascularization and preretinal neovascularization. In order todecrease the risk of intraocular hemorrhage and glaucoma associated withretinal reattachment, a moderate dose of 100 mcg of CAI is givenintravitreally and one-week later successful retinal reattachmentsurgery is performed.

Example 18 Proliferative Sickle Cell Retinopathy

40-year old male with a history of Traction Retinal Detachment andvision loss in one eye from proliferative sickle cell retinopathy. Thepatient developed a Vitreous Hemorrhage in the fellow and has onlyincomplete regression following peripheral scatter laser. Relatively lowdose CAI 1 mg is administered periocularly to cause involution of thesmall amount of residual neovascularization which is causing persistentVitreous Hemorrhage.

Example 19 Choroidal Neovascular Membrane Complications

44-year old white male with a history of Choroidal Neovascular Membranesecondary to high myopia. One eye has Fuch's spots and 20/200 vision.The second eye is affected with a subfoveal Choroidal NeovascularMembrane and 20/80 vision. A single injection of moderate dose, 100 mcg,of CAI is administered intravitreally to hasten neovascular regression.

Example 20 Photodynamic Therapy Complications

32-year old female with multifocal choroiditis and macular scarring inone eye after several sessions of Photodynamic Therapy. She presents inthe second eye with decreased vision and a subfoveal ChoroidalNeovascular Membrane. A mixture of 4 mg of Kenalog, and a high dose of200 mcg of intravitreal CAI are administered to hasten neovascularregression.

Example 21 Treatment of Posterious Infectious Uveitis

35-year old female with posterior infectious uveitis. She presentssymptoms of posterious infectious uveitis due to a parasitic infection.A sustained time-release polymeric implant of 150 to 200 mcg of CAI andof about 0.7 mg dexamethasone is implanted or injected into the insideof the eye.

Example 22 Treatment of Posterious Infectious Uveitis

78-year old male with posterior infectious uveitis and endophthalmitisfollowing four days after cataract surgery. He presents symptoms ofposterious infectious uveitis due to a parasitic infection. A sustainedtime-release polymeric implant of 150 to 200 mcg of CAI with anantibacterial agent or agents and a glucocorticoid is implanted orinjected into the inside of the eye. The sustained time-release implantmay be designed to dissolve.

It should be understood that the embodiments of the invention shown anddescribed in the specification are only preferred embodiments of theinventor who is skilled in the art and are not limiting in any way.Therefore, various changes, modifications, or alterations to theseembodiments may be made or resorted to without departing from the spiritof the invention and the scope of the following claims. While theinvention has been described and illustrated herein by references tovarious specific materials, procedures and examples, it is understoodthat the invention is not restricted to the particular combinations ofmaterial and procedures selected for that purpose. Numerous variationsof such details can be implied as will be appreciated by those skilledin the art. It is intended that the specification and examples beconsidered as exemplary, only, with the true scope and spirit of theinvention being indicated by the following claims. All references,patents, and patent applications referred to in this application areherein incorporated by reference in their entirety.

What is claimed is:
 1. A method of treating uveitis in a patientcomprising administering to the patient a sustained-release systemcomprising a pharmaceutically effective amount of5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide)(CAI).
 2. The method of claim 1, wherein the sustained-release systemcomprises a polymer.
 3. The method of claim 2, wherein the polymerdegrades over time.
 4. The method of claim 3, wherein the polymercomprises polylactic-coglycolic acid (PLGA).
 5. The method of claim 1,wherein the sustained release system comprises a non-erodibleintravetreal implant.
 6. The method of claim 1, wherein the uveitis isposterior infectious uveitis.
 7. The method of claim 6, wherein theposterior infectious uveitis is caused by toxoplasmosis, toxocara orvisceral larva migrans.
 8. The method of claim 1, wherein thesustained-release system further comprises one or more ofanti-inflammatory agent, a steroid and/or an antibacterial agent.
 9. Themethod of claim 8, wherein the steroid is a glucorticoid or fluocinoloneacetonide.
 10. The method of claim 9, wherein the glucocorticoidcomprises dexamethasone.
 11. The method of claim 1, wherein CAI is asonicated microparticle or in a molecular complex in formulation withhydroxypropyl β-cyclodextrin.
 12. A method of treating posteriorinfectious uveitis in a patient comprising administering asustained-release system comprising a pharmaceutically effective amountof5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide)and a steroid to a patient.
 13. The method of claim 12, wherein thesteroid is a glucocorticoid or fluocinolone acetonide.
 14. The method ofclaim 12, wherein the sustained-release system comprises a polymer. 15.The method of claim 14, wherein the polymer comprisespolylactic-coglycolic acid (PLGA).
 16. The method of claim 12, whereinthe steroid comprises dexamethasone.
 17. The method of claim 12, whereinthe sustained-release system further comprises an anti-inflammatoryagent and/or an anti-bacterial agent.
 18. The method of claim 12,wherein the polymer degrades over time.
 19. The method of claim 12,wherein CAI is a sonicated microparticle or in a molecular complex informulation with hydroxypropyl β-cyclodextrin.
 20. The method of claim12, wherein the sustained release system comprises a non-erodibleintravetreal implant.
 21. The method of claim 12, wherein the posteriorinfectious uveitis is caused by toxoplasmosis, toxocara or viscerallarva migrans.